We have established an air pouch-type allergic inflammation model in rats [1,2] and a peritoneal eosinophilia model in rats [3]. Employing the two models, chemical mediators and cytokines responsible for the development of inflammation induced by at allergic mechanisms are investigated to clarify the usefulness of the two models for the screening of anti-allergic inflammatory drugs.

The neurological manifestations of AIDS include dementia, encountered even in the absence of opportunistic superinfection or malignancy. The AIDS Dementia Complex appears to be associated with several neuropathological abnormalities, including astrogliosis and neuronal injury or loss. How can HIV-1 result in neuronal damage if neurons themselves are only rarely, if ever, infected by the vitus\ulcorner In vitro experiments from several different laboratiories have lent support to the existence of HIV- and immune-related toxins. In one recently defined pathway to neuronal injury, HIV-infected macrophages/microglia as well as macrophages activated by HIV-1 envelope protein gp120 appear to secrete excitants/neurotoxins. These substances may include arachidonic acid, platelet-activating factor, free radicals (NO - and O$_2$), glutamate, quinolinate, cysteine, cytokines (TNF-${\alpha}$, IL1-B, IL-6), and as yet unidentified factors emanating from stimulated macrophages and possibly reactive astrocytes. A final common pathway for newonal suscepubility appears to be operative, similar to that observed in stroke, trauma, epilepsy, and several neurodegenerative diseases, including Huntington's disease, Parkinson's disease, and amyotrophic lateral sclerosis. This mechanism involves excessive activation of N-methyl-D-aspartate (NMDA) receptor-operated channels, with resultant excessive influx of Ca$\^$2+/ leading to neuronal damage, and thus offers hope for future pharmacological intervention. This chapter reviews two clinically-tolerated NMDA antagonists, memantine and nitroglycerin; (ⅰ) Memantine is an open-channel blocker of the NMDA-associated ion channel and a close congener of the anti-viral and anti-parkinsonian drug amantadine. Memantine blocks the effects of escalating levels of excitotoxins to a greater degree than lower (piysiological) levels of these excitatory amino acids, thus sparing to some extent normal neuronal function. (ⅱ) Niuoglycerin acts at a redox modulatory site of the NMDA receptor/complex to downregulate its activity. The neuroprotective action of nitroglycerin at this site is mediated by n chemical species related to nitric oxide, but in a higher oxidation state, resulting in transfer of an NO group to a critical cysteine on the NMDA receptor. Because of the clinical safety of these drugs, they have the potential for trials in humans. As the structural basis for redox modulation is further elucidated, it may become possible to design even better redox reactive reagents of chinical value. To this end, redox modulatory sites of NMDA receptors have begun to be characterized at a molecular level using site-directed mutagenesis of recombinant subunits (NMDAR1, NMDAR2A-D). Two types of redox modulation can be distinguished. The first type gives rise to a persistent change in the functional activity of the receptor, and we have identified two cysteine residues on the NMDARI subunit (#744 and #798) that are responsible for this action. A second site, presumably also a cysteine(s) because <1 mM N-ethylmaleimide can block its effect in native neurons, underlies the other, more transient redox action. It appears to be at this, as yet unidentified, site on the NMDA receptor that the NO group acts, at least in recombinant receptors.

Drugs mean not only medicines but also poisons, pesticides, food additives, cosmetics, cleaning agents, environmental pollutants and so on, which are normally considered foreign to the body, It is important to know what happens to these drugs when they get into the body. In the past the metabolic changes of drugs had been referred to as “detoxication mechanism”, but since there are many instances in which drugs are converted in the body to more active substances. Thus, metabolism of drugs is responsible for activation and inactivation of the drugs in the body. The major reactions in drug metabolism are oxidation, reduction, hydrolysis and conjugation. Of these four areas, most of the attention had been focused on the oxidation. Therefore, in contract of ample literatures on drug-oxidizing enzymes, there were relatively few reports on drug-reducing enzymes. In recent years, however, the reduction has received an increasing interest due to its pharmacological or toxicological significance. The present lecture is organized keeping with a focus on drug-reducing enzymes which have been explored by us and by other groups.

In ancient times, respiration's perceived purpose was only to cool the blood, and nasal mucus was thought to be a discharge from the brain. Later it was demonstrated that fluids cannot travel from the brain to the nose. It appears that credit for pointing out the medical significance of bronchial secretions and the biological value of the exocrine function of the lungs belongs to Laennec. He described the “chronic idiopathic pituitous catarrh” known today as bronchorrhea, which is characterized by paroxysms of expectoration. The importance of all this is that airway secretions, and their alterations, became one of the cardinal signs of many respiratory diseases (1).

The development of routine techniques for the isolation and in vitro maintenance of conducting airway epithelial cells in a differentiated state provides an ideal model to study the factors involved in the regulation of the expression of mucocilicary differentiation. Several key factors and conditions have been identified. These factors and conditions include the use of biphasic culture technique to achieve mucociliary differentiation and the use of such stimulators, the thickness of collagen gel substratum, the calcium level, and vitamin A, and such inhibitors, the growth factors EGF and insulin, and steroid hormones, for mucous cell differentiation. Using the defined culture medium, the life cycle of the mucous cell population in vitro was investigated. It was demonstrated that the majority of the mucous cell population in primary cultures is not involved in DNA replication. However, the mucous cell type is capable of self-renewal in culture and this reproduction is vitamin A dependent. furthermore, differentiation from non-mucous cell type to mucous cell type can be demonstrated by adding back a positive regulator such as vitamin A to the “starved” culture. Cell kinetics data suggest that vitamin A-dependent mucous cell differentiation in culture is a DNA replication-independent process and the process is inhibited by TGF-${\beta}$1.

We review the factors which regulate the depth and composition of the human airway surface liquid (ASL). These include secretion from airway submucosal glands, ion and fluid transport across the surface epithelium, goblet cell discharge, surface tension and transepithelial gradients in osmotic and hydrostatic pressure. We describe recent experiments in which we have used low temperature scanning electron microscopy of rapidly frozen specimens to detect changes in depth of ASL in response to submucosal gland stimulation. We also present preliminary data in which X-ray microanalysis of frozen specimens has been used to determine the elemental composition of ASL.

6-Substituted exomethylene 기를 갖는 penam계 화합물이 $\beta$-lactamase 억제제로서 강력한 활성을 보여주고 있어서, 6-exomethylene기 도입에 필요한 1-substituted thioalkyl-1,2,3-triazole-4-carboxaldehyde를 합성하였다. Haloalkanol에 NaN$_3$를 반응시켜서 합성한 azidoalkanol과 propargyl alcohol을 산화시킨 propargyl aldehyde를 반응시켜서 1-(2-hydroxyalkyl-1,2,3-triazole-4-carboxaldehyde을 얻었다. 이것을 trifluoromethanesulfonic anhydride와 triethylamine 존재 하에 mercapto 화합물과 반응시켜서 목적물을 합성하였다.

$\beta$-Lactam계 항생제 내성균의 문제를 해결할 수 있는 $\beta$-lactamase억제제의 개발을 위해서, cephalosporinase에 특히 높은 억제활성을 갖는 새로운 6-exomethylene penamsulfones 화합물을 합성하였다. Dibromopenamsulfone과 thioalkyl triazole-4-carboxaldehyde을 반응시키고, acetic anhydride와 Zn으로 처리하여 E-form과 Z-form의 6-exomethylene penam sulfones을 합성하였다. 이것을 AlCl$_3$으로 deprotection시킨 후, NaOH로 처리하여 6-exomethylene penamsulfone의 Na-salt form으로 목적물질을 합성하였다.

10종의 6-exomethylene penamsulfone 유도체의 Type I, Type IV, TEM 효소에 대한 $\beta$-lactamase 저해 효과를 측정하였다. 7종의 합성 화합물은 Type I penicillinase에 대해서는 저해 효과가 없었으나, Type IV cephalosporinase에 대해서는 기존의 sulbactam이나 tazobactam에 비하여 강한 효과를 나타내었고, 특히 CH 1145는 tazobactam보다 약 30배 강한 활성을 보여주었다. CH 2140은 TEM 효소에 대하여 tazobactam과 유사한 저해효과를 보였다.

새로 합성된 6-exomethylene penamsulfone derivatives 7종에 대해 In Vitro antibacterial activities를 Broth Microdilution법으로 Sulbactam-Na(SAM), Clavulanic acid-K(CLA)와 Cefotaxime-Na(CFT), Ampicillin Trihydrates(AMP), Amoxicillin-Na(AMX), Cefoperazone(CFZ)을 병용투여하여 $\beta$-lactamase 생성 균주(30 여종)에 실시, MIC를 구하였다. 균주에 따라 SAM, CLA에 비해 CH1145, CH1155 등이 우수한 효과를, CH1140, CH2150등이 유사한 효과를 보였다.

Peptide 유도체, 특히 tyrosine을 함유한 peptide 유도체는 항암제 개발을 위한 연구의 관심이 되고 있다. Thiophosphotyrosine을 함유한 peptide는, 종양 발현에 관련되는 여러 효소의 억제제로써, 즉 protein tyrosine kinase(PTK)의 억제제 및 protein tyrosine phosphatase(PTPase)의 억제제 혹은 cytosolic protein의 결합을 방지하는 차단제로 사용할 수 있으며 궁극적으로 항암제 개발에 응용할 수 있다. 이에, t-BOC chemistry를 이용하여 t-BOC-tyrosine을 출발물질로 하고, cyanoethyl 기를 phosphate protecting group으로 사용하여 thiophosphotyrosine을 함유한 peptide 유도체의 합성에 필요한 중요한 중간체 인 N-(tert-butoxycarbonyl)-O-(dicyanoethylthio-phosphene)-L-tyrosine을 합성하였다.

생약(5kg)을 분말로 한 뒤, MeOH 4$\ell$로 24시간, 2회, 환류추출, 추출액을 감압 농축하여 MeOH ex. (450g)을 얻었다. 이것을 물에 현탁시켜 헥산, 벤젠, 에틸아세테이트, 수층으로 분획, L1210 세포에 대하여 이들 분획물과 vincristine의 병용효과를 관찰하였다. 그 결과, 벤젠분획물에서 강한 효과가 있었으므로 컬럼크로마토그라피하여 물질을 분리하였다.

균의 감염에 의해 유도되는 패혈성 shock는 균이 분비하는 elastase가 관여하며, 외부에서 serine pretense inhibitor의 biopolymer의 투여로 균에 의해 유도된 패혈성 shock를 억제시킬 수 있다고 보고되고 있다. 이에 본 연구진은 패혈성 치료제의 개발의 목적으로, 국내에서 민간 약으로 많이 이용되고 있는 백편두로부터 새로운 elastase inhibitor를 분리, 정제하여 부분 아미노산 서열 및 특성을 조사하였기에 발표하고자 한다. 백편두 추출액을 여러 차례에 걸쳐 column chromatography을 수행하면서 얻어지는 각 fraction에 대하여 elastase MCA-기질 및 trypsin MCA-기질을 이용하여 활성 측정 후 elastase 기질 및 trypsin 기질에 대하여 활성을 억제하는 fraction들을 모아 $C_{18}$ open column chromatography 및 $C_{18}$ HPLC 과정을 수행하여 2종류의 trypsin 활성 억제 물질과 1종류의 elastase inhibitor를 분리, 정제하여 각각을 Ti1, Ti2 및 Ti3로 명명하였다. 전기영 동 상에서 단일 hand를 확인한 후 각각의 inhibitor들의 부분 아미노산 서열을 결정하였다.

A series of 6-(N-arylamino)-7-alkylthio-5,8-quinolinedione derivatives(SQ1-12) were newly synthesized for the evaluation of antifungal activities. 6-(N-Arylamino)-7-chloro-5,8-quinolinediones (RCKs) were treated with Na$_2$S/(CH$_3$)$_2$SO$_2$ in EtOH In give SQs. RCKs were prepared by regioselective nucleophilic substitution of 6,7-dichloro-5,8-quinolinediones with arylamines. In the presence of CeCl$_3$, the N-arylamino groups were introduced at the 6-position of 5,8-quinolinedione ring by the regioselective substitution. These derivatives 1-12 were tested for antifungal and also antibacterial activities, in vitro, against Candida sp., Aspergillus niger and Trichophyton mentagrophytes. The MIC values were determined by the two-fold dilution method.

A series of 6-[(N-Halophenyl)amino] -7-Bromo-5,8-quinolinediones (BQs) newly synthesized were tested for antifungal activities, in vitro, against Candida sp., Aspergillus niger and Trichophyton mentagrophytes. The MIC values were determined by the twofold dilution method. Among these derivatives, BQ4 and 7 showed more potent antifungal activities than fluconazole and ketoconazole. BQ 4 and 7 were compared with fluconazole in the treatment of established systemic infections with Candida albicans in normal rats. Intraperitoneally administered BQs for 7 days and 14 days reduced Candida albicans colony count in kidneys and livers as well as fluconazole. The therapeutic potential of BQs has been assessed by evaluating their activities (survival rate) against systemic infections in normal mice with Candida albicans. These compounds, particularly showed activities comparable with fluconazole. BQ4 and 7 had about EO$\_$50/, 0.05, 0.06mg/kg but fluconazole had ED$\_$50/, 6.0mg/kg respectively.

가는잎쇄기풀(Urtica angustifolia Fisch.)은 쇄기풀과(Urticaceae)에 속하는 다년생 초본으로 중약 또는 민간에서 동속 근연식물과 함께 전초를 담마라하여 류마치스성 동통, 산후의 산풍, 소아의 추풍, 경풍,담마진의 치료에 사용되고 있다. Rat에서 실험적 항당뇨 효과를 검토해본 결과 혈당강하 작용이 있는 본 식물로부터 그 혈당강하 성분의 분리에 앞서 식물화학 성분을 규명하고자 본 실험에 착수하였다. 가는잎쇄기풀 전초의 MeOH ex.를 CH$_2$Cl$_2$, EtOAc, n-BuOT 및 $H_2O$로 분획하고 각종 column chromatography를 통하여 다수의 화합물을 분리하였다. 각 화합물은 이화학적 성상 및 spectral data로부터 scopoletin, esculetin dimethyl ether(scoparone), sterol mixture, $\beta$-sitosteryl-3-o-glucoside, kaempferol-3-o-glucoside, quercetin-3-o-glucoside, kaempferol-3-o-rutinoside로 확인하였으며 그 외 다수의 화합물은 그 구조를 규명중이다.

Seeds of Clerodendron trichotomum and Lindera obtusiloba were screened for organic acids. Free organic acids were solid-phase extracted using Chromosorb P from aqueous extract of the seed powder. Organic acids were then converted to tert.-buthldimethylsilyl derivatives with subsequent analysis by dual-capillary gas chromatography and gas chromatography-mass spectrometry. More than twenty organic acids were tentatively identified.

포도당 수송을 증가시키는 것으로 밝혀진 brazilin의 작용이 지방세포의 단백질과 결합하여 나타나는지 알아보기 위한 실험으로 brazilin의 단백결합 성질을 살펴보았다. Brazilin의 단백결합 실험에는 [$^3$H]-brazilin을 사용하여 결합된 양을 방사선 동위원소량으로 확인하였다. 먼저 일반적인 단백결합을 알아보기 위하여 산침전을 이용한 brazilin의 BSA에 대한 결합을 살펴보았다. 그 결과 brazilin은 시간의 경과에 따라 BSA에 결합하는 양이 증가하여 2시간에 최고치에 달했으며 그 양은 약 80% 정도였다. 세포내 단백과의 결합은 BSA에 대한 결합과는 달리 1시간만에 최고치에 달하였다. 세포내 분획에 분포하는 brazilin의 양을 알기 위하여 brazilin처리 후 세포내 분획을 원심분리로 얻어 brazilin의 양을 확인하였다. 그 결과 brazilin은 cytosol에 대부분 존재하며 microsomal membrane과 nuclear에 많은 양이 존재하는 것으로 확인되었다. SDS-PAGE를 이용하여 brazilin과 결합한 각 분획의 단백질을 살펴본 결과 brazilin은 cytosol의 30, 65KD, microsomal membrane의 45, 130KD, nucear의 26, 35KD, plasma membrane의 46, 228KD protein과 반응하는 것으로 확인되었다.

Extract of Pulsatilla Koreana (SB-31) showed promising antitumor activity in vitro (J. Kor Cancer Asso 26:959-963, 1994). We studied the mechanism of cytotoxicity of SB-31. HL-60 cells were cocultivated with various concentrations of SB-31 for 5 hours. The DNAs from HL-60 cells exposed to SB-31 showed the ladder pattern typical of apoptosis. Effect of SB-31 on topoisomerase I activity was determined by slight modification of the method by E. Aflalo(1994). The pBR322 DNA showed dose-dependent increase of R-Form DNA upon incubation with SB-31. The topoisomerase Ⅰ-like activity (Increase of R-Form DNA) was accentuated with higher dose of SB-31. It is postulated that SB-31, which is a fermentation product of Pulsatilla koreana and which loses its activity when kept in ambient temperature for more than 96 hours, may contain topoisomerase Ⅰ-like activity and the enhanced excessive single strand breaks induced by 55-31 may result in apoptosis.

The spinal dorsal horn is the area where primary afferent fibers terminate and cutaneous sensory information is Processed. A number of putative neurotransmitter substances, including excitatory and inhibitory amino acids and peptides, are present in this region and sites and cellular mechanisms of their actions have been a target of numerous studies. In this study, single neurons were acutely isolated and the properties of whole cell current and responses to excitatory and inhibitory neurotransmitters were studied by the patch clamp method. Young rats (7-14 days) were anesthetized with diethyl-ether, and the lumbar spinal cord was excised and cut transversely at a thickness of 30$\mu\textrm{m}$ by Vibroslicer. The treatment of spinal slices with low concentration of proteases (pronase and thermolysin 0.75 mg/$m\ell$) and mechanical dissociation yielded isolated neurons with near intact morphology. Multipolar, ellipsoidal and bipolar, and pyramidal cells were shown. By applying step voltage pulses to neurons held at -70 mV, two types of inward currents and one outward currents observed. The fast activating and inactivating inward current was the Na$\^$+/ current because of its fast kinetics and blocking by 0.5${\mu}$M TTX, a specific blocker of Na$\^$+/ channel. The second type of inward currents were sustained. Based on their kinetics and current-voltage relations, it was likely that the second type of inward current was the voltage-dependent Ca$\^$2+/ current. In the presence of TTX, the steady-state currents mainly represented outward K$\^$+/ current which looked like the delayed rectifier K$\^$+/ current. In addition, the membrane currents produced by agonist of excitatory amino acid (EAA) receptor and the endogenous transmitter candidate L-glutamate were recorded in isolated whole-cell voltage clamped neurons as well as responses to inhibitory amino acids (${\gamma}$-amino butyric acid, glycine). Drugs were applied by a method that allows complete exchange of the solution within 1 sec; an infinite number of solutions can be applied to a single cell.

축산식품중에 잔류하고 있는 sulfamethazine을 검출하기 위하여 sulfamethazine에 대한 단클론항체를 생산하고 이를 이용하여 효소면역측정법을 개발하였다. 면역원은 sulfamethazine에 KLH를 그리고 흡착항원은 BSA를 glutaraldehyde법으로 결합시켰다. 면역원으로 Balb/c mouse를 면역시킨 다음 비장 형질세포률 얻어 myeloma cell과 융합하여 융합잡종세포를 만들었다. Sulfamethazine에 대한 항체를 분비하는 융합잡종세포를 단계회석법과 ELISA를 이용하여 cloning하여 D2, A9, B8, Bl 클론을 얻었다. 이들 클론에서 얻어진 단클론항체를 사용하여 indirect competitive ELISA를 실시하여 표준곡선을 작성하여 본 결과 농도의존성 곡선을 얻을 수 있었다. 4클론중에서 A9 클론을 사용하여 다른 유사한 sulfonamide듣과 p-aminobenzoic acid와 교차반응을 조사한 결과 sulfamerazine에 12.5%의 교차반응을 보였으나 다른 설파제에 대해서는 교차 반응을 보이지 않았다.

Taurine Chloramine (Tau-Cl) inhibits production of nitric oxide(NO) and tumor necrosis factor-alpha (TNF-${\alpha}$) in activated peritoneal macrophages, similar In that previously reported for activated RAW 264.7 cells. In addition, the effect of Tau-Cl and taurine on superoxide anion (O$\_$2/$\^$-/) Production in murine peritoneal exudate polymorphonuclear leukocytes (PMN) was examined. Tau-Cl inhibited O$\_$2/$\^$-/ production in a manner that was dose-dependent and reversible, Taurine also inhibited O$\_$2/$\^$-/ production by stimulated PMN, but at higher concentrations and to a lesser extent than Tau-Cl. The effects of taurine on O$\_$2/$\^$-/ production was attributed to the in vitro formation of Tau-Cl catalyzed by PMN associated halide-dependent myeloperoxidase. In contrast, production of NO and TNF-${\alpha}$ by activated peritoneal exudate macrophages was inhibited by Tau-Cl while taurine was without effect. These data lend support to the notion that Tau-Cl may participate ill the inflammatory responses by modulating production of inflammatory mediators.

Hepatitis B virus (HBV) infection is one of the serious problems in Southeast Asia including Korea because it causes chronic hepatitis, which can easily be transformed In fatal conditions such as cirrhosis and hepatoma. Even though lots of informations on structural characteristics and gene expression mechanisms have been accumulated, the mechanism for HBV-induced hepatocellular injury which is believed to be the consequences of the immunological response is not well understood. In order tn perform immunopathological studies for prevention and treatment of HBV infection, we designed transgenic mice as a disease model which can mimic HBV infection, In this study, a promoter-HBV DNA fragment for the preparation of HBV transgenic mice has been constructed. To add a proper enzyme site on 5' end of HBV gene, total HBV (subtype adr) gene was inserted into BamHI site of pBluescript SK vector and reextracted by PstI-SacI treatment A liver-specific promoter, rat ${\alpha}$ 2u globulin gene promoter, was insrted to pBluescript SK vector and reextracted by BamHI-PstI treatment, Promoter-HBV DNA was constructed by ligation of two fragments using identical PstI sites. For large scale production of promoter-HBV DNA, it was inserted to BamHI-SacI site of pBluescript SK vector.

The involvement of the cyclic AMP (cAMP) effector system in the release of endogenous dopamine and acetylcholine from the rat neostriatum was assessed. Forskolin, an activator of adenylate cyclase, was used to enhance CAMP production, and the consequence of this enhancement on the spontaneous and potassium stimulated release of dopamine and acetylcholine was evaluated. Neostriatal slices were prepared from Fischer 344 rats and after a preincubation period the release of each endogenous neurotransmitter was measured from the same slice preparation. To measure acetylcholine release the slice acetylcholinesterase (AChE) activity was inhibited with physostigmine, but the release from slices with intact AChE activity was also determined (choline, instead of acetylcholine was detected in the medium). Under both conditions forskolin induced a significant dose-dependent increase in the potassium-evoked release of dopamine. In the same tissue preparations the release of neither acetylcholine (AChE inhibited) nor choline (AChE intact) was affected by forskolin. The results indicate that the cAMP second messenger system is involved ill neuronal mechanisms that enhance neuronal dopamine release, but stimulation of this second messenger by forskolin does not further enhance neostriatal acetylcholine release.

Previous studies have shown that kainic acid (KA) causes an elevation of hippocampal proenkephalin mRNA level. However, the role of proto-oncogene products, such as c-Fos, c-Jun and Fra proteins in the regulation of KA-induced proenkephalin mRNA increase in the hippocampus has not been well characterized. Thus, in the present study, the effect of cycloheximide (CHX) on KA-induced proenkephalin mRNA and immediate early gene products induction was examined. After pretreating with either vehicle or CHX (20 mg/kg, s.c.) for 30 min, KA (10 mg/kg) was administered s.c. The animals were sacrificed 1,2, or 8 hrs after KA administration. Total RNA and were isolated for Northern blot assay, and proteins were isolated for Western and electrophoretic gel-shift assays. First, we found that CHX inhibited KA-induced proenkephalin mRNA increase without altering intracellular proenkephalin protein level. Secondly, Western blot assays showed that KA increased c-Fos, c-Jun and Fra proteins at 1,2, and 8 hrs and CHX inhibited these immediate early gene products. Finally, electrophoretic gel shift assays revealed that KA increased both AP-1 and ENKCRE-2 DNA binding activities. Furthermore, CHX attenuated KA-induced AP-1 and ENKCRE-2 DNA binding activities. Both AP-1 and ENKCRE-2 DNA binding activities were abolished by cold AP-1 or ENKCRE-2 oligonucleotides, and further reduced by antibodies against c-Fos or c-Jun. Antibody against CREB reduced ENKCRE-2, but not AP-1, DNA binding activity. Our results suggest that on-going protein synthesis is required for elevation of hippocampal proenkephalin mRNA level induced by KA. All c-Fos, c-Jun, and Fra proteins appears to be involved in the regulation of hippocampal proenkephalin mRNA level induced by KA (This study was supported by a grant from KOSEF).

The (Na, K) ATPase is a membrane ion transporting ATPase composed of an ${\alpha}$ catalytic subunit and a ${\beta}$ glycoprotein subunit. The topology of the rat ${\alpha}$1 and ${\beta}$1 subunits has been studied by insertion of epitope(s) : at the NH2-terminus and COOH-terminus and between Glu117 and Glul18, Lys828 and Arg829, Gln900 and Trp901, and Va1939 and Phe940 of the ${\alpha}$ subunit; and at the NH2-terminus and COOH-terminus and between Glu228 and Tyr229 of the ${\beta}$ subunit. The epitope-tagged ${\alpha}$l, constructs were expressed in HeLa cells to select for stable cell lines expressing a functional (Na, K)ATPase. All constructs, except for the one tagged between Gln900 and Trp901, resulted in ouabain-resistant colonies indicating that modified proteins retained functional integrity. The epitope-tagged ${\beta}$ constructs were transiently expressed in Cos-7 cells. The orientation of the epitopes with respect to the cell membrane was revealed by indirect immunofluorescence performed on permeabilized and non-permeabilized cells expressing the (Na, K)ATPase chains. The results indicate that the ${\alpha}$ subunit has 4 transmembrane segments in the COOH terminal membrane bound domain between residues 760 and 938, and that both the NH2-terminus and the COOH-terminus are in the cytosol; it was not determined whether there are more transmembrane segments between residue 938 and the COOH-terminus. The ${\beta}$ subunit has only one transmembrane spanning region with the NH2-terminus in the cytosol and the COOH-terminus on the extracytoplasmic surface of the plasma membrane.

Annexin I is a member of the annexin family of calcium dependent phospholipid binding proteins and has anti-inflammatory activity by inhibiting phospholipase A$_2$ (PLA$_2$). Recent X-ray crystallographic study of annexin I identified six Ca$\^$2+/ binding bites, which was different types (type II, III) from the well-known EF-hand motif (type I). In this work, the structure of annexin I was studied at atomic level by using $^1$H, $\^$15/N and $\^$l3/C NMR(nuclear magnetic resonance) spectroscopy, and the effect of Ca$\^$2+/ binding on the structure of annexin I was studied, and compared with that of Mg$\^$2+/ binding, When Ca$\^$2+/ was added to annexin I, NMR peak change was occured in high- and low-field regions of $^1$H-NMR spectra. NMR peak change by Ca$\^$2+/ binding was different from that by Mg$\^$2+/ binding. Because annexin I is a larger protein with 35 kDa molecular weight, site-specific (amide-$\^$15/N, carbonyl-$\^$l3/C) labeling technique was also used. We were able to detect methionine, tyrosine and phenylalanine peaks respectively in $\^$13/C-NMR spectra, and each residue was able to be assigned by the method of doubly labeling annexin I with [$\^$13/C] carbonyl-amino acid and [$\^$15/N] amide-amino acid. In $\^$l3/C-NMR spectra of [$\^$13/C] carbonyl-Met labeled annexin I, we observed that methionine residues spatially located near Ca$\^$2+/ binding Sites Were Significantly effected by Ca$\^$2+/ binding. From UV spectroscopic data on the effect of Ca$\^$2+/ binding, we knew that Ca$\^$2+/ binding sites of annexin I have cooperativity in Ca$\^$2+/ binding. The interaction of annexin I with PLA$_2$ also could be detected by using heteronuclear NMR spctroscopy. Consequently, we expect that the anti-inflammatory action mechanism of annexin I may be a specific protein-protein interaction. The residues involved in the interaction with PLA$_2$ can be identified as active site by assigning NMR peaks effected by PLA$_2$ binding.

Bacteriophage T4 endonuclease V initiates the repair of ultraviolet (UV)-induced pyrimidine dimer photoproducts in duplex DNA. The mechanism of DNA strand cleavage involves four sequential stens: linear diffusion along dsDNA, pyrimidine dimer-specific binding,l pyrimidine dimer-DNA glycosylase activity, and Af lyase activity. Although crystal structure is known for this enzyme, solution structure has not been yet known. In order to elucidate the solution structure of this enzyme NMR spectroscopy was used. As a basis for the NMR peak assignment of the protein, HSQC spectrum was obtained on the uniformly $\^$15/N-labeled T4 endonuclease V. Each amide peak of the spectrum were classified according to amino acid spin systems by interpreting the spectrum of $\^$15/N amino acid-specific labeled T4 endonuclease V. The assignment was mainly obtained from three-dimensional NMR spectra such as 3D NOESY-HMQC, 3D TOCSY-HMQC. These experiments were carried out will uniformly $\^$15/N-labeled sample. In order to assign tile resonance of backbon atom, triple-resonance theree-dimensional NMR experiments were also performed using double labeled($\^$15/N$\^$13/C) sample. 3D HNCA, HN(CO)CA, HNCO, HN(CA)HA spectra were recorded for this purpose. The results of assignments were used to interpret the interaction of this enzyme with DNA. HSQC spectrum was obtained for T4 endonuclease V with specific $\^$15/N-labeled amino acids that have been known for important residue in catalysis. By comparing the spectrum of enzyme*DNA complex with that of the enzyme, we could confirm the important role of some residues of Thr, Arg, Tyr in activity. The results of assignments were also used to predict the secondary structure by chemical shift index (CSI).

Nitric Oxide Synthase(NO synthase: EC.1.14.13.39)는 생체내에서 L-arginine을 기질로 하여 nitric oxide(NO)와 L-citrulline의 생성을 매개하는 효소로서 뇌, 간장, 신장, 체장등 대부분의 주요장기와 근육세포, 신경세포 등 거의 모든 조직에 분포하고 있다. NO synthase에 의해 생성되는 NO는 혈관이완작용, 신경전달 물질로서의 작용, 면역 담당세포에서의 세포 독작용 등 많은 생리현상에 중요한 역할을 하는 것으로 알려져 있다. 특히 체장에서는 췌외분비 기능의 항진에 있어 세포내 cGMP level의 변동이 NO와 연관된다는 사실에 주목하고 있으며 본연구실에서도 이에 관한 연구가 진행중이다. 따라서 본 연구에서는 소 췌조직의 100,000$\times$g cytosol을 효소원으로 하여 다음과 같이 NO synthase의 분리, 정제를 시행하였다. Ammonium sulfate로 30%(176g solid ammonium sulfate/$\ell$) 포화, 침전 후 2',5'-ADP agarose 및 calmodulin-agarose affinity chromatography를 연속적으로 시행하여 NO synthase를 분리하였으며 electrophoresis상에서 약 160kd의 분자량을 나타내었다.

Nitric oxide synthase(NO Synthase:E.C.1.14.13.39)는 생체내에서 L-arginine을 기질로 하여 citrulline과 nitric oxide(NO)를 생성하는 효소로서, 최근 연구에 의하면 2/3 부분 간 절제술 후 prereplicative phase동안에 발현되는 것으로 알려져 있다. 한편, 생체내에서 NO Synthase에 상경적 길항제인 methylarginine에 관해서도 수년간 많은 연구가 진행되어 왔다. 이들의 생성 기전은 protein methylation에 의해 생성된 methylated protein이 생체 내에서 분해되어 생성된다고 알려져 있으나, 정확한 기전에 대해서는 아직 논란의 여지가 많다. 따라서, 본 연구에서는 In vivo 실험을 통해 부분 간 절제술 후 시간대별로 간 조직에서의 NO Synthase 활성도와 혈청에서 NO의 최종 대사물인 Nitrite/Nitrate를 측정하였으며, 또한 NO Synthase 조절에 관여하는 세포내 기질인 arginine과 억제 인자인 methylarginine함량을 간 조직 및 혈청에서 측정하고, 세포 신호 전달체계에 관여하는 cyclic GMP 함량을 측정함으로써, 부분 간 절제술 후 간 재생동안에 NO Synthase 활성도와 methylarginine 및 arginine과의 상관 관계를 규명하고, 간 재생동안, 생성된 nitric oxide의 역할을 연구하려한다.

Streptococcus pneumoniae (pneumococcus), the most common cause of bacterial pneumonia, has an ample polysaccharide(PS) capsule that is highly antigenic and is the source of PS vaccine. This investigation was undertaken to optimize the culture conditions for the production of capsular PS by type 1 pneumococcus. Among several culture media, brain heart infusion (BHI) and Casitone based media were found to support luxuriant growth of pneumococcus type 1 at the same level. Because BHI medium is rather expensive and more complex than the Casitone based media, the Casitone based media was used to study optimization of the culture condition. The phase of growth which accomodated maximum PS production was logarithmic phase. Concentrations of glucose greater than 0.2% did not enhance growth or PS production. Substitution of nitrogen sources with other resources or supplemention of various concentrations of metal ion (with the exception of calcium ion) had adverse effects on growth and PS production. On the other hand, low level aeration was beneficial for increased PS production. Addition of 3 mg/I concentration of methionine, phenylalanine, and threonine were found to enhance growth and PS production. The synergistic effect of all the favorable conditions observed in pneumococcal growth assays provided a two-fold cumulative increase in capsular PS production.

세포질에 존재하는 100 kDa Phospholipase $A_2$(cPLA$_2$)는 인지질의 sn-2 위치의 에스테르결합을 가수분해함으로서 Prostaglandin과 Leukotriene등 Eicosanoids 생합성의 전구체인 아라키돈산과 Platelet activating factor(PAF)를 생합성하는 전구체를 동시에 생성시키는 효소로 염증과 세포손상등에 중요한 역할이 기대된다. 본 효소의 활성화 기전을 규명하고자 하는 최근의 활발한 연구에도 불구하고 불명확한 점이 많은 것이 현실이다. 특히 세포를 자극하였을 때 유리되는 아라키돈산의 증가율과 세포를 파괴한 후 조제한 가용성분획에서 측정한 활성의 증가율과는 많은 차이를 나타냈다. 이러한 결과부터 cPLA$_2$ 효소 자체를 활성화시키는 어떤 인자를 가정하였다. 최근, PLA$_2$의 또다른 형태인 14 kDa의 분비성 PLA$_2$의 in vitro 활성을 증가시키는 인자가 동정되어 그 생화학적 특성이 규명되고 있으나 이 인자는 cPLA$_2$의 활성에는 아무런 증가효과를 나타내지 않았다. 본 연구자들은 소의 뇌조직에서 cPLA$_2$의 활성을 증가시키는 인자를 발견하고 그의 생화학적인 특성을 규명하였다. 돼지 비장에서 정제한 cPLA$_2$를 사용하였으며 소의 뇌 조직의 가용성 분획으로부터 본 활성화 인자를 동정하였으며 그 활성분획을 양이온 크로마토그라피로서 Mono S EPLC와 Superose 12 Sepharose gel filtration 크로마토그라피를 이용하여 더욱 분리한 결과 약 70 kDa과 25 kDa에서 각각 용출되었다. 이렇게 부분정제한 활성은 췌장에서 분리한 group I과 흰주의 group I과 흰주의 혈소판에서 분리한 group II PLA$_2$에 대해서는 아무런 증가효과를 나타내지 않는 반면, cPLA$_2$의 활성만을 약 5배 증가시켰다. 본 활성은 cPLA$_2$ 효소량의 증가에 따라 활성의 증가효과가 정차 감소하므로 화학량적인 반응(Stoichiometric reaction)일 것으로 예상되었다.

In order to gain insight into the mechanism of the regulation of cytochrome P450IAl by arylhydrocarbon, the 5'-flanking region of a trout CYP450IAl 5'flanking DNA was cloned into pCAT-basic vector and it was transfected into Hepa-1 cells. 3MC treatment to hepa Ⅰ cells transfected with fish CYP450IAl-CAT construct results in mRNA increased by 2.81 fold when it was compared with that of control This increase of mRNA was decreased by concomitantly treated flavonoids such as morin. The levels of CAT mRNA that was treated with morin was 29.2-58.0% of 3MC stimulated CAT mRNA. Further investigation to find out if there are DRE, XRE or negative regulatory cis element in CYP450IA1 gene was undertaken. Results of the deletion study of 5'flanking DNA of trout P450IA indicate the existance of the negative(-1600 ～ -1300). CAT mRNA was about two-fold higher in deleted trout CYP450IAl-CAT construct transfected cells compared to the wi Id type trout CYP450IAl-CAT construct transfected cells. And The stimulatory effect of 3MC was no longer observed in col Is containing deleted CAT construct. [Supported by grants from the Korean Ministry of Education]

BQ-123, a selective $ET_{A}$ receptor antagonist, reverses various responses induced by Endothelin-1 and it has been reported that BQ-123 ameliorates the cerebrovascular constriction, hypertension, and decrease of blood flow. Previously, we announced that the level of Endothelin-2 increase in the brain and spinal cord of EAE-induced lewis rat and showed the origin of ET-1 is activated macrophages. Intracisternal injection of $ET_{A}$ receptor antagonist, BQ-123(10nmol) was done for visualizing the role of endothelin-1 on the pathogenesis of EAE. BQ-123 apparently blocked the severity of clinical score of EAE and decreased the histologically observed inflammatory region. The blocking effect on the progression of EAE model following BQ-123, suggests that BQ-123 is a physiological antagonist in terms of development of the sign of multiple sclerosis.

Endothelin has $ET_{A}$ type and $ET_{B}$ type receptors, and it has been thought that ET-1 proves vasoconstriction effect via $ET_{A}$ receptor and vasodilation via $ET_{B}$ receptor. Recently, it has been reported that $ET_{B}$ receptor is also related to the vaso-constriction. Bosentan is a $ET_{A+B}$ receptor antagonist, and proves it's effect on trauma and ischemia. We already announced that the level of Endothelin-1 increase in the brain and spinal cord of EAE-induced lewis rat and showed the origin of ET-1 is activated macrophages. Intracisternal injection of Bosentan, $ET_{A+B}$ receptor antagonist, (300nmol/body) was done for observing the role of endothelin-1 on the pathogenesis of EAE. Bosentan ameliorated the severity of clinical score of EAE and decreased the histologically observed inflammatory region. The blocking effect on the progression of EAE model suggests that Bosentan is a physiological antagonist in terms of development of the sign of multiple sclerosis.

Endothelin-1, which is a peptide originally isolated from cultured porcine aortic cell, has been found to play a crucial role in potentiating the vasoconstriction mitogenesis, and chemotaxis. In the present study, we examined the level of endothelin-1 in the brain, spinal cord and blood from rat with experimental allergic encephalomyelitis(EAE). At the peak stage of EAE(grade 3), endothelin-1 level in the spinal cord of rat with EAE increased two folds as compared with that of sham-treated rats, and subsequently decreased to the level of control at the recovery stage. In the endothelin-1 immunocytochemistry, endothelin-1 immunopositive cells and ED-1, macrophage marker immunopositive cells observed in the spinal cord of peak stage(grade 3). These Findings suggest that endothelin-1 play the important role in progression of EAE.

Pathogenesis of experimental allergic encephalomyelitis was involved with infection, inflammatory reaction, immune reaction etc. We studied on relation of blood vessel system and EAE. So, we measured blood pressure, heart rate and relaxation of isolated blood vessel in control and EAE-induced lewis rats. Blood pressure was decreased before EAE clinical sign (0-20day), but was increased from 23day. In isolated blood vessel, acetylcholine-treated relaxation was different on control, maximum EAE stage, recovery stage. Acetylcholine-treated relaxation was reduced 30% in recovery stage. but, sodium nitroprusside had similar relaxation reaction.

우루소데옥시콜린산을 1% CMC에 현탁하여 25mg/kg 의 농도로 각각 1, 3, 6, 10일간 1일 3회씩 투여한 후. 고정틀에 묶어 고정시키고 4시간 동안 스트레스를 가하였다. 스트레스 부과후 복부를 개복하여 혈액을 채취하고 스트레스지표인 ALP, AST, ALT, LDH를 측정하였다. 한편 일부 혈액은 백혈구, 림프구수를 측정하기 위해 이용되었다. 부신 및 비장을 적출하여 무게를 측정하고 부신은 분쇄하여 아스코르빈산 함량을 정량하였다. 스트레스 부과시간에 따른 백혈구, 림프구수의 변화실험은 별도로 시행하였다.

Artemisia extract powder (DA-9601)의 일반약리 작용을 mice, rat 및 rabbit에서 검토하였다. DA-9601은 rota-rod test에 의한 운동 협조능, 초산 writhing 반응, tail-flick test 및 randall-sellito test를 이용한 진통작용, thiopental sodium의 수면작용, 장관 수송능, carrageenan의 염증, 현수에 의한 근이완작용, 정상체온, strychnine의 경련작용, 수정관, 자궁각 및 Evans blue의 피부혈관 투과성에 별다른 영향을 미치지 않았다. DA-9601은 토끼의 적출회장 운동을 억제하였고 혈압 강하작용과 이뇨작용을 나타내었다

Helicobacter pylori (H. pylori) is a microaerophilic spiral bacterium and infection by it in the human stomach causes gastritis, furthermore, it is considered to be involved in the pathogenesis of peptic ulcers and the development of gastric carcinoma. We assessed the inhibitory activity of new antiulcer drugs against Helicobacter pylori. The activities of new antiulcer agents against Helicobacter pylori strains were determined by the standard agar dilution method with blood agar base #2, supplemented with 5% sheep blood and 4 antibiotics to support growth of these organisms. They were inoculated by multipoint inoculator and incubated at 37$^{\circ}C$ for 3 days under microaerophilic atmosphere. The MIC of antiulcer agents was the lowest concentration that inhibited visible growth of these organisms. According to results of various biochemical tests, these bacteria were identified as Helicobacter pylori strains. And the MIC results showed that the strains were very susceptible to omeprazole and YJA20379s. Some of YJA20379s were more potent than omeprazole. These results suggest that our new antiulcer drugs have potent inhibitory activity against Helicobacter pylori, so that our new antiulcer drugs might be useful for the clinical eradication of gastrointestinal Helicobacter pylori.

Eupatilin은 급성위궤양 모델인 HCl-EtOH model에서 0.3mg/kg에서 50% 억제하였으며 10mg/kg에서 95% 최대효과를 나타내었고, indomethacin model에서는 0.8mg/kg에서 50% 억제하였으며, 10mg/kg에서 97% 최대억제효과를 나타내었다. Cyclooxygenase-1의 활성은 1.7, 5.8$\mu\textrm{g}$/$m\ell$에서 prostaglandin E$_2$와 prostacyclin의 생성을 각각 2배 촉진시켰으나, cyclooxygenase-2에 대한 활성에는 영향을 주지 않았다. FMLP로 활성화시킨 호중구에 대한 활성은 0.6$\mu\textrm{g}$/kg에서 50% 억제하여 oxygen free radical 소거활성을 나타내었고, 반면 xanthine oxidase 및 iron-dependant lipid peroxidation 활성에 대해서는 78.8, 26.9$\mu\textrm{g}$/$m\ell$에서50% 억제를 보였다. 5-lipooxygenase 활성은 2.1$\mu\textrm{g}$/$m\ell$ 에서 염증매개인자인 leukotriene B$_4$ 생성을 50% 억제하였다. 이상의 결과로부터 애엽추출의 항궤양 효과는 prostaglandin의 생성촉진 및 oxygen free radical 과 leukotriene B4와 같은 공격인자의 생성억제 등의 복합적인 작용에 기인하는 것으로 생각된다.

급성 간질환모델에 대한 인진호 추출분획의 간장 약효검색, 방법 1. $CCl_4$ 간장해 : SD계 수컷 흰쥐에 $CCl_4$와 olive oil 혼합액(1:4v/v%)을 체중100g당 0.2$m\ell$씩 복강내 투여하였으며, 시험약물은 $CCl_4$ 혼합액 투여 4시간전 및 6시간후에 경구로 2회 투여하였다. 48시간후에 부검하여 혈청을 얻어 간기능검사 항목인 ALT 및 AST 활성을 측정하였다. 2. D-Galactosamine 간염 : SD계 수컷 흰쥐에 D-GalactosamineㆍHCl을 650mg/kg씩 복강내 투여하였으며, 시험약물은 $CCl_4$ 간장해시험과 동일하게 2회 경구투여하였다. D-Galactosamine 투여 24시간 후에 부검하여 혈청을 얻어 간기능검사 항목인 ALT 및 AST 활성을 측정하였다. 3. 담즙울체모델 : SD계 수컷 흰쥐에 ANIT 100mg을 olive oil 1$m\ell$에 현탁시켜 80$m\ell$/kg b.wt. 용량으로 1회 경구투여하였으며, 시험약물은 ANIT 투여전 2시간, 투여 후 6, 22, 28시간 간격으로 4회 경구투여하였다. ANIT 투여 47시간 후에 1시간 동안 담즙을 채취하였고, ANIT 투여 48시간째 채혈하여 혈청내 총빌리루빈치를 측정하였으며 담즙배출량, 담즙중 담즙산량도 측정하였다. 4. 약물투여 음성대조 : 1% CMC-Na 용액(10$m\ell$/kg b.wt.) 양성대조 : Silymarin(25mg/kg), UDCA(25mg/kg), DDB(37.5mg/kg) 인진호추출분획 : 인진호 원료의 수침액인 BE분획의 수득률을 기준으로 하여, KP(180mg/kg), PS-1 및 PS-2(300mg/kg), EE(500mg/kg), HH(640mg/kg), BE(1500mg/kg)

간재생 및 만성 간질환모델에 대한 인진호 추출분획의 간장 약효검색 방법 1. 간세포 재생능력에 대한 효과 : SD계 수컷 흰쥐(체중 300-350g)를 마취하에서 복부정중선을 따라 개복후 간의 중앙엽 및 좌측엽을 절제하여 봉합하고 수술조작을 끝낸 추 4시간, 익일부터 7일간 오후 5시에 약물을 경구투여하고 마지막투여 후 20시간 절식, 부검하여 간재생률 및 BSP 정체율을 측정하였다. 2. $CCl_4$ 만성간장해 모델에 대한 효자 SD계 수컷 흰쥐(체중 200-250g)에 $CCl_4$ : olive oil 40%용액을 0.5 $m\ell$/kg b.wt. 용량으로 6일간 결구투여 한 후 7일째부터 매일 시험약물을 $CCl_4$ 혼합액 투여 4시간전 및 4시간후에 2회 경구투여하였으며 이 조작을 8일간 하고 마지막 투여 24시간 후에 부검하여 혈액내 ALT 및 AST치, 간조직내 total protein, triglyceride 및 hydroxyproline 양을 측정하였으며, 간중량도 측정하였다. 3. 약물투여 음성대조 : 1% CMC-Na 용액(10$m\ell$/kg b.wt.) 양성대조 : silymarin(25mg/kg), DDB(37.5mg/kg) 인진호추출분획 : KP(180mg/kg), PS-1(300mg/kg), PS-2(300mg/kg), EE (500mg/kg ), HH(640mg/kg), BE(1500mg/kg)

천연물 성분의 약효와 간독성에 관여하는 기전연구의 일환으로 사염화탄소로 유발된 흰쥐에서 혈청 중의 효소 활성도, 지질함량 및 과산화지질을 측정하여 홍화자 추출물과 분획물이 사염화탄소로 유발되는 간독성에 미치는 효과를 규명하고자 하였다. 홍화자는 홍화(Carthamus tinctorius L.)의 씨로서 성미는 신, 온하며 활혈거제 및 소 지통의 효능을 가진 구어혈제로서 자궁에 대한 흥분작용과 관상동맥을 확장시키는 작용이 있으며 혈압을 하강시키는 작용이 있다. 또한 고cholesterol 혈증을 낮추고 혈전폐색성 맥간염을 치료하는 효과도 있다고 한다.

The antihistaminic action of eighteen herbal medicines was investigated by the radioligand binding and functional assays. The hexane fractions of Trichosanthis radix, Mori cortex radicis and Evodiae fructus dose-dependently inhibited [$^3$H]mepyramine binding to H$_1$ receptor and histamine-induced contraction in guinea-pig brain homogenates and isolated guinea-pig ilea, respectively. Antihistaminic action of the hexane and ethylacetate fractions of Mori cortex radicis and the hexane fraction of Evodiae fructus was more potent than their antimuscarinic action evaluated from the inhibition of [$^3$H]QNB binding and carbachol response. The ethylacetate and chloroform fractions and six known flavonoids from Scutellariae radix also inhibited histamine-induced contraction, but antihistaminic potencies of these fractions and compounds were almost identical with their antimuscarinic potencies. The hexane fractions of Mori cortex radicis and Evodiae fructus, as shown in ketotifen, inhibited selectively the increase of cutaneous vascular permeability induced by histamine. However, wogonin (SC-1) from Scutellariae radix was a nonselective inhibitor for the effect of histamine and serotonin on the vascular permeability. These results demonstrate that the hexane and ethylacetate fractions of Mori cortex radicis and the hexane fraction of Evodiae fructus have the selective histamine H$_1$ receptor blocking activities.

It has been known that pinusolide isolated from Biota orientalis inhibits specifically [$^3$H] PAF binding to and PAF-induced aggregation of rabbit platelets in vitro. The present study was undertaken to evaluate the effects of pinusolide on hypotension and bronchoconstriction due to PAF. PAF(i.v.) in doses of 0.01 to 0.3 $\mu\textrm{g}$/kg induced a dose-dependent hypotension without tachyphylaxis in rat. Pinusolide(5～20 mg/kg, i.v.) inhibited PAF(0.03 $\mu\textrm{g}$/kg, i.v.)-induced hypotension dose-dependently, while it failed to block the hypotensive action of histamine, serotonin and acetylcholine. It inhibited bronchoconstriction and cutaneous vascular permeability induced by PAF in anesthetized guinea-pigs and rats, respectively, but it showed no inhibitory effect on the increase in bronchial resistance by histamine and acetylcholine. These results strongly suggest that in vivo pinusolide acts as a selective antagonist of PAF.

모체의 당뇨병은 신생아 호흡부전 증후군의 중요한 소인의 하나이며, 이것은 태아의 폐성숙이 지연되어 폐포를 안정화시키는 표면활성물질의 결여에 기인하는 것으로 알려져 있다. 폐의 표면활성물질은 지질과 소량의 단백으로 이루어진 복합물질로서 그 주성분은 phosphatidyl choline이며 동물의 종속에 관계없이 임신 말기에 그 양이 현저히 증가한다. 양수내 인지질은 주로 태아 폐에서 유래하므로 임신 말기의 양수내 lecithin/sphingomyelin(L/S)비 측정으로 태아의 호흡부전 증후군을 예측할 수 있다. 일반적으로 당뇨병 모체의 양수는 L/S비가 낮아 당뇨병 산모로 부터 출생한 신생아에서 호흡부전증의 발생빈도가 높으므로 이를 예방하기 위하여 부신피질호르몬의 투여가 고려되고 있다.

본 실험은 간장허혈 및 재관류시 야기되는 간장 손상에 대해 vitamin C와 E 각각의 효과와 이들의 병용효과를 알아보고자 하였다. 실험군은 흰쥐에 vitamin E(25mg/kg)를 실험전 3일간 투여한 군, vitamin C(100mg/kg)를 실험 5분전 경정맥주사한 군 및 vitamin C와 E의 병용 투여군등의 3군으로 하여 각각에 허혈을 유발시킨 후 (60분) 재관류 1시간, 5시간에 간세포 손상정도(AI.T, AST, liver wet-weight to dry-weight ratio), 지질과산화(MDA), 담즙분비변동(bile flow, bilirubin, cholate output) 및 약물대사효소계의 변동(cytochrome P$_{450}$, aminopyrine-N-demethylase, aniline p-hydroxylase activity) 등을 관찰하였다. 실험결과로는 허혈 및 재관류로 인한 ALT, AST MDA는 재관류 5시간에 최고치를 이루었으며 이는 vitamin C와 vitamin E의 각각 투여로 억제되었고, 특히 vitamin C와 E의 병용투여로 더욱 현저하게 억제되었다. 간세포 부종의 지표인 liver wet-weight to dry-weight ratio도 vitamin C와 E의 병용투어로 유의성있게 억제되었다. 담즙분비량 및 담즙산량은 vitamin C 투여와 vitamin C와 E 병용투여로 허혈 및 재관류로 감소된 양을 증가시켰고, 특히 vitamin C와 E의 병용투여는 담즙분비량에 있어 현저한 상승을 나타내었다. 허혈 및 재관류로 인한 cytochrome P$_{450}$양의 감소와 aminopyrine N-demethylase 활성의 억제는 vitamin C 투여와 vitamin C와 E의 병용투여에 의해 유의성 있게 증가하였다. 이상의 결과로 보아 vitamin C와 vitamin E는 각각 허혈 및 재관류로 인한 간장손상을 완화시켰으며 특히 vitamin C와 E의 병용투여는 상승적으로 적용하여 간세포손상을 더욱 억제시킴을 알 수 있었다.

The C-terminus ends of the second putative transmembrane domains of both m1 and m2 muscarinic receptors contain a triplet of amino acid residues consisting of leucine (L), tyrosine (Y) and threonine (T), This triplet is repeated as LYT-LYT in m2 receptors at the interface between the second transmembrane domain and the first extracellular loop. Interestingly, however, it is repeated in a transposed fashion (LYT-TYL) in the sequence of m1 receptors. In this work we employed site-directed mutagenesis to investigate the possible significance of this unique sequence diversity for determining the distinct differential drug-receptor interaction and cellular function at m1 muscarinic receptor. Mutation of the LYTTYL sequence of m1 receptors to the corresponding m2 receptor LYTLYT sequence, however, did not result in a significant change in the binding affinity of the agonist carbachol or in the affinity of the majority of a series of receptor antagonists which are able to discriminate between wild-type m1 and m2 receptors. Surprisingly, the LYTLYT ml receptor mutant demonstrated markedly enhanced coupling to activation of phospholipase C without a change in its coupling to increased cyclic AMP formation. There was also an enhanced receptor sensitivity in transducing elevation of intracellular Ca$\^$2+/. These changes were not due to alterations in the rate of receptor. desensitization or sequestration, On the other hand, the reverse LYTLYT-LYTTYL mutation in the m2 receptor did not alter its coupling to inhibition of adenylate cyclase, but slightly enhanced its coupling to stimulation of PI hydrolysis, Our data suggest that the LYTTYL/LYTLYT sequence difference between ml and n12 muscarinic receptors is not involved in determining receptor pharmacology. On the other hand, while these differences might play a role in the modulation of muscarinic receptor coupling to PI hydrolysis, they are not important for specifying coupling of various subtypes of muscarinic receptors to different cellular signaling pathways.

Through molecular cloning, five muscarinic receptors have been identified. The muscarinic receptors can be generally grouped according to their coupling to either stimulation of phospholipase C (m1, m3, and m5) or the inhibition of adenylate cyclase (m2 and m4). Each m1, m3, and m5 receptors has the additional potential to couple to the activation of phospholipase A$_2$, C, and D, tyrosine kinase, and the mobilization of Ca$\^$2+/. However, the differences in coupling efficiencies to different second messenger systems between these receptors have not been studied well. Ectopic expression of each of these receptors in mammalian cells has provided the opportunity to evaluate the signal transduction of each in some detail. In this work we compared the coupling efficiencies of the m1, m3 and m5 muscarinic receptors expressed in chinese hamster ovary (CHO) cells to the Ca$\^$2+/ mobilization and the stimulation of neuronal nitric oxide synthase (nNOS). Because G protein/PLC/PI turnover/[(Ca$\^$2+/])i/NOS pathway was supposed as a main pathway for the production of nitric oxide via muscarinic receptors, we studied on ml, m3 and m5 receptors. Stimulation of guanylate cyclase activity in detector neuroblastoma cells was used as an index of generation nitric oxide (NO) in CHO cells. The agonist carbachol increased the cGMP formation and the intracellular [Ca$\^$2+/] in concentration dependent manner in three types of receptors and the increased cGMP formation was significantly attenuated by scavenger of NO or inhibitor of NOS. m5 receptors was most efficiently coupled to stimulation of nNOS, And, the coupling efficiencies to the stimulation of neuronal nitric oxide synthase in three types of receptors were parallel with them to the Ca$\^$2+/ mobilization.

In this work we investigated coupling of the m2 and m4 subtypes of muscarinic acetylcholine receptors expressed in chinese hamster ovary (CHO) cells to activation of neuronal nitric oxide synthase (nNOS). Stimulation of guanylate cyclase activity in detector neuroblastoma cells was used as an index of generation of nitric oxide (NO) in CHO cells. The agonist carbachol induced marked time and concentration-dependent enhancement of the activity of nNOS at m2 receptors. In sharp contrast, the response in CHO cells transfected with the m4 receptor gene was similar in magnitude to that observed in non-transfected cells, suggesting lack of significant coupling of m4 muscarinic receptors to NO signaling. This novel observation of functional divergence of the two muscarinic receptor subtypes at the level of activation of nNOS is quite intriguing, in light of the currently accepted dogma that they belong to the same functional class. This functional selectivity was not due to differential effects on intracellular Ca$\^$2+/ concentration, since activation of both subtypes of muscarinic receptors produced a comparable, albeit quite small, Ca$\^$2+/ signal. Taken together, our present data strongly suggest that the generally assumed functional equivalence of m2 and m4 muscarinic receptors should be carefully reexamined. These data also suggest the presence of alternate mechanisms of activation of nNOS, which might be operative in the absence of large changes in the concentration of cellular Ca$\^$2+/. The latter mechanisms are expected to be activated by m2, but not m4 muscarinic receptors. Both sets of findings are quits important in regards to refining the functional classification of muscarinic receptor subtypes and the cellular mechanisms of activation of NOS.

Muscarinic acetylcholine receptors contain two highly conserved tyrosine residues which are located within or at the extracellular border of the second transmembrane domain. These tyrosine residues are located at positions 82 and 85 of the sequence of the ml subtype of muscarinic receptors. In this wok, we studied the involvement of these two residues in ligand binding to and agonist-induced activation this receptor subtype. our data suggest an important role for these two tyrosines in these processes, with a more prominent role for the tyrosine residue located at position 82 than that located at position 85. Evidence is also provided that while the aromatic moiety of these tyrosine residues is important for antagonist binding, both this moiety and the tyrosine phenolic hydroxyl group are involved in agonist binding and receptor activation.

Changs in the glutamatergic nervous system following AF64A injection into lateral ventricle were studied in rats. Rats were treated with the infusion of AF64A (3mM) into lateral ventricle At a week after the infusion of AF64A into lateral ventricle, rats were sacrified and each brain resions was dissected ; striatum, hippocampus and frontal cortex. At these resions, total glutamate and other amino acids levels. [$^3$H]Mk801 binding sites and glutamine synthetase activity were measured using HPLC-ECD, ligand binding assay and enzyme activity assay, respectively. The levels of total glutamate were decreased in striatum, hippocampus and frontal cortex Also, the levels of total glycine and taurine were decreased in all examined regions. Furthermore, the levels of total aspartate and GABA were decreased in both hippocampus and frontal cortex but these didn't alter in striatum. Additionally, the levels of total glutamine were decreased in both striatum and frontal cortex. The u\numbers of [$^3$H]MK801 binding sites were differently dffected in each brain resions ; the decrease in striatum, the increase in frontal cortex and no change in hippocampus Glutamine synthetase activity in striatum was significantly decreased. But, that in both hippocampus and frontal cortex didn't alter These results suggest that changes in the glutamatergic nervous system in three regions are induced by following AF64A injection into lateral ventricle in rats.

Changes in glutamate release and uptake on cerebellar cells after the chronic exposure to lead were investigated. Rats were received 0.25% lead acetate in drinking water from the beginning of the pregnancy. The control group was given 0.125% sodium acetate in drinking water. The cerebellar cells from 7 or 8 day-old pups were cultured. Amino acid release from cerebellar granule cells and the glutamate uptake into cerebellar glial cells were measured using HPLC-ECD. Basal glutamate release and NMDA-induced glutamate release didn't show significant difference. However, the other amino acids in the granule cells obtained from lead exposed pups were less released than the control after the stimulation by NMDA (50$\mu$M). SNAP-induced (50$\mu$M) glutamate release was significantly reduced in granule cells prepared from lead exposed pups. The basal glutamate uptake in glial cells didn't show any difference. However, the uptake in glial cells prepared from lead exposed pups was significantly less blocked by PDC (24$\mu$M) compared to the control group. These results indicate that lead exposure to the mother might affect the Excitatory amino acid system during the development of the offspring.

Effects of ginseng total saponin(GTS) on changes in the glutamatergic nervous system induced by AF64A were studied in rats. Rats were pretreated with the infusion of AF64A (3mM) into lateral ventricle and were posttreated with GTS (50mg/kg, j.p) for 1 week. Twenty four hrs after the last administration, rats were sacrified and each brain resions was dissected ; striatum, hippocampus and frontal cortex. At each brain regions, total glutamate and other amino acids levels, [$^3$H]MK801 binding sites and glutamine synthetase activity were measured using HPLC-ECD, ligand binding assay and enzyme activity assay, respectively. The AF64-induced increase in the levels of total glutamate in hippocampus were significantly decreased by the administration of GTS. Furthermore, that compared with saline and GTS was decreased in striatum. The levels of total GABA compared with saline and GTS were declined in frontal cortex. Moreover, the AF64A-induced decrease in the levels of total taurine were significantly increased by the administration of GTS to extents of normal states. The numbers of [$^3$H]MK801 binding sites were differently affected in brain resiojns ; the decrease in hippocampus and no change in both striatum and frontal cortex, Glutamine synthetase activity was significantly increased in hippocampus. In comparision with saline and GTS, that was significantly decreased in striatum These results suggest that GTS may adjust the levels of glutamate, GABA and taurine constantly and may induce increase of glutamine synthetase activity declined.

The effects of the anti-tumor agent, SDZ-y2434, on rat kidney were investigated to predict the toxicities of its derivatives and to develope less toxic derivatives. After adjusted in metabolic cages for 5 days, rats were treated SDZ-62434(acute : 25mg/kg, i.p, once and 50mg/kg, i.p., once; subacute ; 10mg/kg, i.p., daily for 7 days). Kidney weights and urine volume during the treatment were observed. Creatinine concentration, protein concentration and the activities of N-acetyl-${\beta}$-D-glucosaminidase (NAG), alanine aminopeptidase (AAP), ${\gamma}$-glutamyl transpeptidase (GGT) and lactate dehydrogenase(LDH) in 24 hr urine were also determined. The kidney weights after the acute and subacute administration didn't show any difference. Urine volume increased 5 days after the acute administration (50mg/kg) and 3 days after the subacute administration. The excretion of creatinine was increased 5 days after the acute (50mg/kg) and subacute administration. However, the protein excretion didn't show any change. NAG acivity declined 7 days after the subacute administration. AAP and GGT activites increased 3 days after the acute administration (50mg/kg) but, returned to the control value. LDH activity showed continuousely high value after the subacute administration. These results indicates that the acute administration of SDZ-62434 might damage on glomerulus and that the subacute administration might be cytotoxic to kidney cells.

Intraperitoneal injection of morphine (5 mg/kg) in mice every other day for 8 days produced conditioned place preference (CPP). CPP effects were evaluated by assessing the difference in time spent in the drug-paired compartment and the saline-paired compartment of the place conditioning apparatus. The injection of a non-competitive NMDA antagonist, MK-801 (0.05 and 0.1 mg/kg, i.p.), prior to and during morphine treatment in mice Inhibited morphine-induced CPP. The development of postsynaptic dopamine (DA) receptor supersensitivity in mice displaying a morphine-induced CPP was evidenced by the enhanced response in ambulatory activity to the DA agonist, apomorphine (2 mg/kg). MK-801 inhibited that development of postsynaptic DA receptor supersensitivity MK-801 also inhibited apomorphine-induced climbing behavior, suggesting that MK-801 Inhibits dopaminergic activation mediated via the NMDA receptor.

The present study examined chronic effects of transient focal cerebral ischemia on the substantia nigra, a remote exofocal area, using immunohistochenmical and receptor autoradiographic techniques. Transient focal cerebral ischemia was induced by middle cerebral artery (MCA) occlusion for 60 or 90 min followed by reperfusion using silicone-coated 4-0 nylon monofilament in male Wistar rats. After 1- or 2-week reperfusion following transient MCA occlusion, there were partial losses of tyrosine hydroxylase-immunoreactive dopaminergic neurons, incieases in glial fibrillary acidic protein-immunoreactive cells (gliosis), decreases in [$^3$H]YM-09151-2 binding for dopamine D$_2$ receptors, and marked atrophy in the ipsilateral substantia nigra. The precise mechanism(s) of exofocal damage to the substantia nigra is remained to be elucidated.

The purpose of the present study was to determine whether in vivo noradrenergic neural activity in the locus coeruleus is related to the development of hypertension. Two groups of animals were prepared, 1) young spontaneously hypertensive rats (SHR) and 2) age-matched normotensive wistar kyoto rats (WKY). At il weeks of age, the release of norepinephrine (NE) and 3,4-dihydroxyphenylglycol (DOPEG) from locus coeruleus of young SHR and WKY as an index of neural activity were determined by in vivo microdialysis along with blood pressure (BP) at three conditions : 1) normal; 2) elevated BP by systemic injection of phenylephrine and 3) alpha-1 adrenoceptor stimulated by perfusion of phenylephrine into the locus coeruleus through microdialysis probe. Basal releases of NE and DOPEG from the iocus coeruleus were 0.415+/-0.089pg/20min, 1.311+/-0.293 pg/20min in SHR and 0.204+/-0.078 pg/20min, 1.492+/-0.365 pg/20min in WKY respectively. Basal release of NE from the locus coeruleus of SHR was significantly greater than that of WKY. Phenylephrine systemic injection caused elevation of BP in both SHR and WKY in a dose related manner. Following phenyephrine injection, the releases of NE and DOPEG from the locus coeruleus of SHR were significantly decreased, whereas there were no significant changes in the releases of NE and DOPEG In young WKY. Alpha-1 adrenoceptor stimulation in the locus coeruleus by perfused phenylephrine through microdialysis probe caused pressor responses in both SHR and WKY, but the magnitude of pressor response in SHR was larger compared with that in WKY. The result from the present study suggests that noradrenergic neural activity in locus coeruleus may contribute as one of triggering factors for the expression of hypertension in young SHR.

The putative role of vasoactive intestinal polypeptide (VIP) as non-adrenergic non-cholinergic (NANC) neurotransmitter has been studied in rabbit corpus cavernosum. In the presence of atropine and guanethidine the short and prolonged electrical field stimulation (EFS, 2～16 ㎐) induced a frequency-dependent relaxation which was abolished by tetrodotoxin (0.3 ${\mu}$M), a nerve conductance blocker. The neurogenic relaxant reponses were not affected in the presence of VIP-inactivating peptidase, ${\alpha}$-chymotrypsin (2 units/$m\ell$), whereas VIP-induced relaxation were completely abolished. Inhibition of nitric oxide synthase by N$\^$G/-nitro-L-arginine (10～100 ${\mu}$M) caused concentration-dependent inhibition to the neurogenic relaxant responses and at 100 ${\mu}$M the relaxations were virtually abolished. In contrast NO (3～30 ${\mu}$M) and VIP (0.001～l ${\mu}$M)-induced relaxation were unaffected. The inhibitory effect of L-NNA was reversed in the presence of L-arginine (5 mM), the precursor of the NO biosynthesis. Hemog1obin (20～60 ${\mu}$M), sequestering NO in the extracellular space, abolished the NO-evoked relaxation and also caused a concentration-dependent inhibition to the neurogenic relaxation. These observation indicate that NANC relaxation induced by prolonged EFS of rabbit corpus cavernosum is also mediated mainly by nitric oxide as same as that of short EFS, and suggest that VIP is not involved in NANC relaxation of rabbit corpus cavernosum and NO would not be produced by VIP in this tissue.

Brazilin은 포도당 수송을 증가시키는 물질로 autooxidation에 의하여 산화되면서 hydrogen peroxide를 생성할 것으로 추정되었으며, hydrogen peroxide는 phosphatase를 억제하여 포도당 수송을 증가시키는 것으로 보고되었다. 따라서 본 실험에서 brazilin의 산화에 의한 hydrogen peroxide의 생성여부를 확인하고 phosphatase 활성에 미치는 braziline의 작용을 살펴보았다. 먼저 UV absorption spectra를 이용하여 brazilin이 반응액중에서 구조적인 변화를 일으키는지 확인하였다. Hydrogen peroxide의 생성은 rhodamine 123를 이용한 형광측정법으로 측정하였으며, phosphatase의 활성은 pH에 따른 phosphatase의 활성을 p-NPP의 탈인산화의 UV 흡광도 변화로 측정하였다. PP2A의 활성은 phosphorylase a를 기질로 하여 측정하였다.

Mucin release from hamster tracheal surface epithelial(HTSE) cells can be stimulated by extracellular ATP via activation of P$_2$ purinoceptors located on the cell surface which appears to be coupled to phospholipase C via G proteins. However, our preliminary data indicate that the ATP-induced mucin release involves, in part, activation of PKC, but not an increase in the intracellular Ca++ level, suggesting the presence of another pathway which is separate from the PLC-PKC pathway, In this study, we intended to confirm the previous observation and subsequently identify an additional mechanism. Confluent HTSE cells were metabolically labeled with either $^3$H-glucosamine or $^3$H-arachidonic acid(AA), and release of either $^3$H-mucin or $^3$H-AA was quantified following various treatments. $^3$H-mucin was assayed using the sepharose CL-4B gel-filtration method, whereas $^3$H-AA liberation was measured by counting $^3$H-radioactivity in the chase medium. We found that: (1)Desensitization of PKC by pretreatment with PMA completely abolished the mucin releasing effect of PMA but partially inhibited the ATP-induced mucin release; (2) ATP increases release of $^3$H-AA in a dose-dependent fashion; (3) mepacrine, an inhibitor of PLA$_2$, attenuates ATP-induced mucin release in a dose-dependent fashion. These results confirm our previous notion that the PLC-PKC pathway is responsible, in part, for ATP-induced mucin release. Furthermore, activation of PLA$_2$ appears to be an additional pathway which is involved in ATP-induced mucin release.

The objectives of this study were to evaluate the pharmacokinetics of SJ-151 which is a new NSAID in male Beagle dogs following a single oral dosing. Seven beagle dogs (10-l2kg) were all administered a single oral gavage(10mg/kg) of SJ-151 tablet and serial blood samples of approximately 5$m\ell$ were then drawn from the cephalic vein of each animal at 0(predose), 0.25, 0.5, 1, 1.5, 2, 3, 4, 6, 8, 12 and 24hours postdosc. SJ-151 (Cinmetacinㆍbutendiol)and cimetacin were quantified in the plasma fraction by HPLC assay. The pharmacokinetic parameters calculated from the plasma concentrations of SJ-151 in dayl are Cmax(ng/$m\ell$)509$\pm$248, Tmax(hr) 1.50$\pm$0.45, AUC(ng.hr/$m\ell$) 1,750$\pm$762, tl/2$\alpha$ 0.98$\pm$0.30, t1/2$\beta$ 12.0$\pm$6.75. The pharmacokinetic parameters calculated from the plasma concentrations of Cinmetacin in day 1 are Cmax(ng/$m\ell$)258$\pm$74.1, Tmax(hr)2.42$\pm$ 0.92, AUC(ng.hr/$m\ell$) 1,820$\pm$318, t1/2$\alpha$ 1.74$\pm$0.49, t1/2$\beta$ 25.4$\pm$13.4.

(중략) 1)에탄올 0.08g/kg 급성투여군은 선조체 5-HT, DOPAC, HVA, DOPAC/DA, HVA/DA ratio를 현저히 증가시켰다. 반면, 3g/kg 급성투여군은 도파민 및 세로토닌 신경활성에 전혀 영향을 미치지 않았다. 2) 에탄을 3g/kg을 21회 연속투여군은 선조체 DA 및 5-HT와 5-HIAA 함량을 유의성있게 증가하였으나 도파민 대사산물에는 영향을 주지 않았다. 3) 35주령 흰쥐는 에탄을 3g/kg 급성투여군에서 5-HT함량을 감소시켰으나 3g/kg 21회 연속투여군은 5-HT함량을 유의성있게 증가시켰다. 4) 7주령과 35주령 모두 21회 연속투여에서 DA 및 5-HT함량을 현저히 증가시켰다.

In order to elucidate the correlation between the lipid peroxidation and hepatotoxicity, the formation of malondialdehyde (MDA) in liver homogenate and serum, and the transaminase activities were determined in intoxicated by ascorbic acid-Fe$\^$2+/-ADP in rat. In a model of ascorbic acid-Fe$\^$2+/-ADP hepatotoxicity, G009, which was obtained from Ganoderma lucidum IY009, exhibited anti-lipid peroxidative effect in rat liver homogenate, and that MDA values of the liver homogenate decreased from 48.1% to 74.8% in comparision to controls (p<0.01) Also, the MDA formation in serum inhibited 66.5% at 100 mg/kg of G009. Serum levels of glutamic oxaloacetic transaminase(GOT) and glutamic pyruvic transaminase(GPT) in peroxidizer-induced rats treated with G009 was decreased compared with control. Especially, The formation of lipid peroxides in serum was related to GPT levels. These results that G009 has a protective effect on ascorbic acid-Fe$\^$2+/-ADP-induced hepatic injury through an inhibition of lipid peroxidation in liver.

Effects of Panax ginseng were tested on morphine-and steroids-induced immunosuppression, focusing on mechanism and identification of active components. To investigate overall effects of morphine and ginseng total saponin (GTS) on immune system, body weight and lymphoid organ weight were measured. Morphine significantly reduced body weight, spleen/body weight, and thymus/body weight ratio. GTS, at 100mg/kg (oral), restored spleen/body weight ratio. Because morphine is known to increase corticosterone level, serum corticosterone level was measured by radioimmunoassay. Serum corticosterone was increased by morphine and it was restored to the control level by GTS 100mg/kg (oral). In vitro proliferation studies were also conducted to study the effects of ginseng on steroids-induced immunosuppression. While ginsenoside Rg$_1$ and ginseng alkaloids were effective on proliferation and dexamethasone-induced death of thymocytes, 50% ginseng ethanol extract and polysaccharides were effective on splenocytes. In vivo mprohine-induced apoptosis of thymus was partly protected by GTS.

홍삼 총 사포닌 투여군은 대조군과 비교시 total free radical 및 malondialdchydc 농도는 유의상 있게 감소되었으며, 단백질의 carbonyl 농도는 다소 감소하는 경향을 나타내었다. 그리고 홍삼 총 사포닌 투여군의 경우 Cu, Zn-SOD, catalasc, GSII reductase 등의 항산화 효소와 nonprotein-SH가 대조군 보다 증가되었다. 홍삼 총 사포닌의 구성성분들인 ginsenoside Rb$_1$, Rb$_2$, Rc, Rd, Re, Rg$_1$, Rh$_1$, Rh$_2$, Rf 중 ginsenoside Rh$_2$는 catalase 활성을 대조군보다 유의성있게 증가시켰으며, ginsenoside Rh$_1$ 및 Rc의 경우 GSII peroxidase 활성이 증가하는 경향을 나타내었다. 그리고 Cu, Zn-SOD의 경우 ginsenoside Rc는 대조군보다 유의성있게 감소시켰으며, GSII reductase의 경우 유의성있는 변화는 관찰되지 않았다.

1, 흰쥐에서 IPM 단독투여시보다 DHP-I 저해제인 CS와의 병용투여시 AUC가 4배 이상, t$_{1}$2/가 약 3배 증가하였으며 MEPM의 경우는 AUC가 5배 이상, t$_{1}$2/가 약 1.8배 증가하였다. 2. Guinea pig에서 IPM 단독투여시보다 CS와의 병용투여시 AUC가 약 2.5배, t$_{1}$2/가 약 1.4배 증가하였으며 MEPM의 경우는 AUC, t$_{1}$2/ 모두 거의 변함이 없었다. 3. 위의 결과 흰쥐에서 IPM과 MEPM에 대한 CS의 영향이 매우 큰 것으로 나타났으며 따라서 약물동력학적인 파라메타에도 커다란 변화를 주었다. 반대로 Guinea Pig에서는 CS의 병용투여로 IPM의 AUC에만 변동을 주었을 뿐 t$_{l}$ 2/에는 크게 영향을 미치지 않았고 MEPM의 체내동태에는 거의 영향을 미치지 않았다. 4. Guinea pig에서 전체 투여량에 대한 MEPM의 요중회수율은 66.58$\pm$3.44%이었고 IPM의 경우는 7.00$\pm$0.95%이었다. 그러나 DHP-I 저해제인 CS와의 병용투여시 IPM의 요중회수율이 57.77$\pm$11.46%로 증가하여 MEPM과 거의 비슷한 양상을 나타냈다. 이는 guinea pig에서 DHP-I에 의한 IPM의 대사가 CS에 의해 차단됨을 시사하는 것이라고 사료되며 guinea pig에 존재하는 DHP-I은 MEPM보다 IPM에 대해 더 높은 친화도를 갖는 것으로 사료된다. 5. Carbapenem계 항생제의 약물동력학적 스크리닝을 위한 소동물로는 흰쥐보다도 guinea pig가 적합한 것으로 생각되나 동물종차에 의한 약물동력학적 연구 및 DHP-I의 활성에 대한 연구가 더 면밀히 이루어져야 할 것으로 생각되며 BO-2727, panipenem, biapenem 등 다른 carbapenem 항생제에 대한 광범위한 약물동력학적 연구가 진행되어 새로운 carbapenem계 항생제 개발에 따른 인체내 거동을 예측할 수 있는 치적의 실험동물모델이 확립되어야 할 것이다.

The 116 annual meeting of the Pharmaceutical Society of Japan was held in Kanazawa from March 27 to 29 in 1996, Our presentation will be concerned to a few topics in this meeting. Recently not only pharmaco- and toxico-kinetic studies but also pharmaco- and toxico-dynamic studies are beginning to be worthy of notice. This is quite reasonable, because the blood concentration-time curves of a parent compound must be useful, but it is no more than a cross section for pharmaco- and toxico-dynamic in many organs and tissues as main reaction site for drug metabolism,

Paraquat(1.1'-dimethyl-4,4'-bipyridinium ion, PQ)는 전세계적으로 가장 많이 사용되고 있는 농약으로 자살 또는 실수로 마시는 경우 예외없이 폐독성으로 사망하게 된다. 하지만 아직까지 임상에 사용되어지고 있는 효과적인 독성 경감제는 전무한 실정이다. 현재까지 밝혀진 paraquat의 주된 독성기전은 NADPH Cytochrome P$_{450}$ reductase에 의해 산화,환원 반응을 거치는 동안 free radical을 생성하여 세포막에 지질과산화를 일으켜서 세포막의 기능상실과 cell death를 일으킨다고 보고되고 있다. 따라서 본 연구에서는 항산화작용이 뛰어난 아미노산인 taurine(TA)의 radical scarvenging 효과에 의한 PQ 독성경감효과를 in vivo에서 검색하였고, in vitro에서 TA의 PQ 독성경감 mechanism을 밝히고자 하였다. in vivo에서 PQ의 간독성(s-GOT,s-GPT), 신장독성(BUN, Creatinine), 폐 및 전신독성(ALP, MDA, G-6-phosphatase)의 정도를 혈액 및 조직균질액 중에서 검색함으로써 TA의 독성억제효과를 측정하였다.

DA-1131은 gram positive bacteria와 Pseudomonas aeruginosa를 포함한 gram negative bacteria에 대하여 광범위한 항균 spectrum과 우수한 항균력을 나타내었다. 각종 임상분리균주에 대한 항균력 시험 결과, gram positive bacteria 중 methicillin-resistant Staphylococcus aureus(MRSA)에 대하여는 DA-1131이 가장 우수한 항균력을 나타내었으며, methicillin-susceptible S. aureus(MSSA)에 대하여는 MEPM, CPR 및 CAZ보다 약 2-50배의 우수한 항균력을 나타내었으나 IPM/CS보다는 동등이하의 항균력을 나타내었다. Gram negative bacteria에 대하여는 IPM/CS, CAZ 및 CPR보다 우수한 항균력을 나타내어 0.2 $\mu\textrm{g}$/$m\ell$ 이하의 농도에서 91%의 Serratia marcescens, 89%의 Proteus mirabilis, 76%의 Morganella morganii 및 시험에 사용된 Enterobacteriaceae에 속하는 전균주의 생육이 억제되었다. P. aeruginosa 에 대하여 DA-1131은 1.56 $\mu\textrm{g}$/$m\ell$ 이하의 농도로 시험균주 전체의 생육을 저해하였으며, MEPM의 약 2배, IPM/CS의 약 4배의 강한 항균력을 나타내었다. CAZ에 내성인 Enterobacteriaceae 임상분리균주에 대한 DA-1131의 항균력은 CAZ 감수성균주에 대한 항균력과 동일한 것으로 나타났다.

배지의 영향에 대하여 검토하기 위하여 SDA, MHA, NA 및 BHIA를 사용하였으며, 시험균주 중 Pseudomonas aeruginosa에 대하여만 NA에서의 MIC 값이 낮게 나타났을 뿐 나머지 균주에 대하여는 배지에 따른 항균력 변화가 관찰되지 않았으며, 배지의 pH를 5.4-8.4까지 변화시켰을 때 pH 변화에 따른 DA-1131의 유의적인 항균력 변화와 접종균량에 따른 항균력 변화가 관찰되지 않았다. Methicillin-resistant Staphylococcus aureus(MRSA)에 대하여는 DA-1131과 vancomycin 간의 상승 및 상가작용이 관찰되었고, P. aeruginosa, Klebsiella pneumoniae, Enterobacter cloacae 및 Serratia marcescens 등의 균종에 대하여는 ciprofloxacin의 상가작용이 확인되었다. Sisomicin은 P. aeruginosa에 대한 상승 및 상가작용이 인정되었으나 ceftazidime은 DA-1131에 의한 길항작용이 관찰되었다.

DA-5018은 여러 opiate수용체 실험에서 morphine 또는 naloxone 보다 10-100배 정도 낮은 친화력을 나타내었다. 기니픽 회장표본과 랫드 수정관표본의 전기자극 실험에서는 DA-5018의 의해 유도된 수축반응이 naloxone의 영향을 받지 앉았고, 랫드에서의 진통효과도 naloxone 전처치에 의해 차단되지 않았으므로, DA-5018은 opiate 수용체를 경유하지 앉는 것으로 생각된다. 또한, OA-5018 120$\mu$M에서 cyclooxygenase 생성을 50% 증가시켰고 mM 농도에서 5-lipooxygenase 합성을 약간 억제하였으므로, NSAID계 유사약물이 아님을 확인하였다. DA-5018은 기니픽 기관 표본에 대해 capsaicin과 동일하게 수축반응을 나타내었고, 이것은 capsazepine 전처리에 의해 억제되었다. 척수에서의 substance P 유리활성은 capsaicin보다 약 9배 강한 것으로 나타났다. 또한 랫드에 DA-5018 진통유효용량인 1mg/kg을 피하주사한 후 분리한 척수에서 capsaicin에 의한 substance P의 유리활성은 15분 후에 감소하였고 120분 후에는 회복되었다. 따라서 OA-5018의 진통작용에도 substance P의 고갈이 관여하는 것으로 생각된다. 이상의 결과로부터, DA-5018의 진통작용은 capsaicin수용체를 매개하는 것으로 사료된다.

1. 초산 writhing법에서 DA-5018, morphine, capsaicin 및 acetaminophen의 E $D_{50}$ 은 각각 0.1, 0.3, 1.4 및 45.4 mg/kg이었으며, formalin법에서 DA-5018과 morphine의 E $D_{50}$은 0.17과 4.3 mg/kg이었다. Randall-Selitto법에서 DA-5018, morphine, capsaicin 및 acetaminophen의 E $D_{50}$은 0.66, 3.9, 4.2 및 643 mg/kg이었다. Tail flick법에서 DA-5018, morphine 및 capsaicin의 E $D_{50}$ 은 2.0, 2.6 및 14.2 mg/kg이었고 hot plate법에서 DA-5018과 capsaicin의 E $D_{50}$은 1.7 및 26.5mg/kg이었다. 그러나 열자극에 의한 동통모델에서 acetaminophen은 진통효과가 없었다. 2. Streptozotocin유발 당뇨랫드에 DA-5018 0.2, 0.5 및 1.0mg/kg 또는 capsaicin 10 mg/kg 투여시 1, 3, 7 일째에서 각각 유의성있는 동통역치의 증가를 나타내었으나 ketoprofen(10 mg/kg) 및 desipramin(10 mg/kg) 투여시는 동통역치의 유의성있는 증가가 나타내지 않았다. 관절염 동통모델에서 DA-5018, ketoprofen 및 capsaicin을 투여한 후 7일째에 동통역치를 2배 증가시키는 용량은 각각 0.66, 3.76 및 17.38 mg/kg이었다. 3. 이상의 결과로부터 DA-5018은 morphine 및 capsaicin에 비해 동등 이상의 효력을 갖고 있으며, 비스테로이드성 진통제보다 효능이 우수함을 확인하였다.

DA-1131은 IPM이 D2 channel 특이적인 세포외막투과성을 나타내는 것과는 달리 D2 channel 이외에 azthreonam 투과경로도 이용하는 것으로 확인되었다. IPM 감수성 및 내성인 P aeruginosa 균주들에 대하여 $\beta$-lactamase의 inducible activity, hydrolytic susceptibility, affinity를 측정한 결과, Inducible activity는 DA-1131, PPM 및 MEPM이 거의 동일하였으며, 3가지 약물 모두 가수분해에 대한 안정성을 나타내었다. 그러나 $\beta$-lactamase에 대한 affinity는 IPM이 가장 컸고, MEPM, DA-1131의 순으로 감소하였다. DA-1131은 P8P2와 PBP3를 동시에 저해하며, IPM은 PBP2의 저해 후 순차적으로 PBP3를 저해하였고 이러한 시험결과는 PBPs blinding pattern과 밀접한 관계가 있는 것으로 알려진 균형태변화로도 확인되었다.

DA-1131, imipenem(IPM) 및 meropenem(MEPM)은 각종 $\beta$-lactamase를 산생하는 세균에 대하여 우수한 항균력을 나타내었으나 cefpirome(CPR), ceftazidime(CAZ) 및 azthreonam(AZT)의 경우에는 extended broad spectrum cephalosporinase 산생 균주를 포함하여 일부 균주의 내성획득이 확인되었다. DA-ll3l의 $\beta$-lactamase Inducible activity는 DA-1131, IPM 및 MEPM이 거의 동일하였으며, imipenemase 산생균주로 동점된 Serratia marcescens 11001이 산생하는 $\beta$-lactamase이외의 효소에는 대부분 가수분해되지 않는 결과를 나타내었다. S. marcescens 11001이 산생하는 $\beta$-lactamase에 대한 효소역학상수는 DA-1131, IPM 및 MEPM에서 모두 유사하였고, $\beta$-lactamase에 대한 affinity도 큰 차이를 나타내지 않았다.

각종 동물 및 인체 신장 유래 DHP-I에 의한 DA-1131, imipenem(IPM) 및 meropenem(MEPM)의 속도 상수를 측정한 결과, DA-1131은 동물종에 관계없이 가장 안정성이 큰 결과를 나타내었고, 인체 DHP-I에 대한 Vmax/Km 값이 IPM의 21.9%로 관찰되어 IPM보다 하위 기질인 것으로 확인되었다. DA-1131, IPM/CS 및 MEPM/CS의 20mg/kg투여 후의 혈중농도 반감기(T$_{1}$2/)는 각각 11.4분, 8.9분, 10.3분이었으며, 1$\mu\textrm{g}$/$m\ell$ 이상의 혈중농도룰 유지하는 시간은 66.6 분, 55.9 분, 63.1 분이었다. DA-1131, DA-l131/CS, IPM/CS, MEPM 및 MEPM/CS의 40 mg/kg 투여 후 24시간 동안의 뇨중 배설율은 57.9 %, 61.3%, 22.6 %, 11.3% 및 65.9%이었으며, 각 약물을 40 mg/kg 투여 15분 후 DA-ll3l의 폐중 농도는 11.2$\mu\textrm{g}$/g으로 DA-l131/CS, IPM/CS 및 MEPM/CS와 비슷한 결과를 나타내었으며 T/P ratio도 DA-1131, DA-l131/CS, IPM/CS 와 MEPM/CS 투여군에서 거의 동일한 것으로 확인되었다. 신장중 농도는 DA-1131 과 DA-l131/CS의 경우 29.l$\mu\textrm{g}$/g 및 34.2$\mu\textrm{g}$/g으로 큰 차이를 나타내지 않았으나, IPM/CS, MEPM 및 MEPM/CS 투여군에 비하여는 높은 결과로 나타났고 T/P ratio도 DA-1131과 DA-l131/CS 투여군이 IPM/CS, MEPM 및 MEPM/CS 투여군보다 놓은 것으로 확인되었다.

DA-1131은 Staphylococcus aureus, Escherichia coli, Klebsiella pneumoniae, Enterobacter cloacae, Proteus mirabilis, Proteus vulgaris 및 Pseudomonas aeruginosa 의 시험균주에 대하여 1-4의 MBC/MIC 비를 나타내었으며, 약물첨가에 의한 세균증식억제 과도 매우 우수하였으므로 광범위의 강력한 살균력을 나타내는 것으로 확인되었다. DA-1131의 PAE는 S. aureus Smith에 대하여는 1.28시간, K. pneumoniae 1에 대하여는 0.65시간, P. aeruginosa 93에 대하여는 1.90시간으로 나타나, MIC 이하의 낮은 농도에서도 세균에 대한 생육저해 효과가 관찰되었으며, 이러한 현상은 E. coli K-12에 DA-1131을 1/4 MIC(0.0125 $\mu\textrm{g}$/$m\ell$) 농도로 작용시켰을 때 균의 팽화와 신장 및 구형화가 동시에 일어나는 등 균형태변화를 초래하는 효과와 큰 상관성을 나타내었다. 또한, DA-1131은 감염방어효 에 매우 큰 영향을 끼치는 mouse macrophage와 매우 우수한 협력적살균작용을 나타내어 E. coli K-12 생세포는 1/16 MIC 이상의 DA-1131 공존하에서 쉽게 식균소화(phagocytosis) 되었다.

Staphylococcus aureus, Klebsiella pneumoniae, Enterobacter cloacae, Proteus mirabilis 및 Pseudomonas aeruginosa에 의한 mouse 복강내감염증에 대하여 DA-1131은 IPM/CS와 MEPM/CS에 비하여 현저히 우수한 치료효과를 나타내었다. 백혈구감소 mouse에서의 P. aeruginosa에 의한 복강내감염증에 대하여도 DA-1131은 IPM/CS와 MEPM 및 MEPM/CS보다 동등 이상의 우수한 치료효과를 나타내었다. K. pneumoniae를 감염균으로 한 mouse 호흡기감염 후 생존율 평가 시험에서 DA-1131 투여 mouse는 IPM/CS 및 MEPM/CS에 비하여 현저히 우수한 생존율을 나타내었고, 동시에 MEPM/CS와 유사한 폐내 생균수의 감소 pattern이 확인되었으며, IPM/CS에 비하여 균의 재증식을 억제하는 효과가 우수하였다. P. mirabilis에 의한 mouse 요로감염증에서는 DA-1131 투여군의 신장내 생균수감소는 MEPM/CS와 동등한 수준인 것으로 나타났으나 IPM/CS에 비하여는 매우 우수하였다.

새로운 항암성 백금(II)착제 SA: ([Pt( trans-1-dach) (DPPE)]ㆍ2NO$_3$, SB: [Pt (cis-dach)(DPPP)]ㆍ2NO$_3$, SC: [Pt(cis-dach)(DPPE)]ㆍ2NO$_3$에 대한 일반 약리작용이 rat, mouse 및 rabbit에서 검토되었다. Rota-rod test에 의한 운동 협조능, thiopental-Na의 수면 작용, 현수에 의한 근이완 작용, 정상체온, strychnine 유발 경련, 담즙분비, 수정관, 자궁각, 피부혈관 투과성, 국소마취, 용혈 및 심전도에 대하여 SA, SB. SC는 별다른 영향을 미치지 않았단. 적출회장과 십이지장에 대하여 SA는 수축반응을 나타내었으나 SB와 SC는 하등의 영향이 없었으며 혈액 응고에 대하여 SA는 prothrombine time을 단축시켰으나 SB, SC는 별다른 영향이 없었고 호흡에 대해서는 SA, SB, SC 다같이 호흡수를 억제하였다. 적출심장 기능에 대하여 SA, SB, SC 다같이 고농도에서 억제적으로 작용하였다.

The influence of glucocorticoids on the secretory responses of catecholamines (CA) evoked by acetylcholine (ACh), DMPP, McN-A-343, excess K$\^$+/ and Bay-K-8644 from the isolated perfused rat adrenal gland and to clarify the mechanism of its action. The perfusion of the synthetic glucocorticoid dexamethasone (10-100 uM) into an adrenal vein for 20min produced relatively a dose-dependent inhibition in CA secretion evoked by ACh (5.32mM), excess K$\^$+/ (56mM), DMPP (a selective nicotinic receptor agonist, 100uM for 2min), McN-A-343 (a muscarinic receptor agonist, 100uM for 4min), Bay-K-8644 (a calcium channel activator, 10 uM for 4min) and cyclopiazonic acid (a releaser of intracellular Ca$\^$2+/, 10uM for 4min).

백금착물 항암제 후보물질로서 합성된 KBP31705-Cl27, KBP30603-901을 기존 화합물인 cisplatin 및 carboplatin과 pharmacokinetic profile을 비교 검토하였다. 웅성 Sprague-Dawley rat을 sodium pentobarbital 마취하에서 방광 및 대퇴동맥과 정맥에 polyethylene tubing을 사용하여 catheterization시켰다. Urine 배출이 안정화되었을 때 대퇴정맥내로 cisplatin과 KBP31705-Cl27은 2 mg/kg, carboplatin과 KBP 30603-901은 20mg/kg 용량으로 주사한 후 2, 4, 6, 8, 10, 15, 20, 30, 45, 60, 120 분에 대퇴동맥으로부터 혈액을 채취하였으며 urine은 약물투여 후 1시간 간격으로 4시간동안 채취하였다. Plasma와 urine중의 platinum농도는 inductively coupled plasma-mass spectrometer를 사용하여 측정하였고, pharmacokinetic parameters는 non-linear least square computer Program인 PCNONLIN을 이용하여 산출되었다. 혈중 platinum농도와 시간의 관계에서 KBP31705-Cl27은 cisplatin과 비교하여 alpha-phase에서 혈중 농도는 낮지만 비슷한 양상을 나타내었고, beta-phase에서는 비교적 느리게 소실됨을 보였다. Urine으로 4시간동안 배출된 platinum양은 각각 투여량의 52, 56%로서 두 약물에서 비슷하였다. 또한 KBP30603-901은 carboplatin과 비교하여 alpha-phase는 거의 비슷한 패턴을 나타내었으나, beta-phase는 훨씬 느리게 감소하여 반감기가 길다는 것을 보여 주었으며 또한 이것은 urine으로 4시간동안 배출된 platinum양이 KBP30603-901의 경우 투여량의 46%로서 carboplatin의 59%보다 적게 배출된다는 data와도 일관됨을 보여주었다. 이상의 결과로 볼 때 KBP30603-901이 다른 백금착물 항암제보다 체내에 머무르는 시간이 길어 혈중에서 보다 오랜 시간동안 머물러 높은 유효농도를 유지할 수 있을 것으로 사료된다.

In previous studies we reported that the levels of RBC, hemoglobulin and hematocrit in SAM Rl and SAM P6 were increased significantly from 7 day after oral administration of the pilose antler extract, 5g/kg/day, and were lasted during the study. Therefore, this study was performed to elucidate mechanism of erythropoietic action by the extract administration. SAM Rl and SAM P6 were chosen as experimental animals. At age of 12 weeks, pilose antler extract were given 0.3 and 5 g/kg/day (p.o.) each for 0, 7 and 14 days in both animals. Complete blood cells (CBC) such as WBC, lymphocytes, monocytes, granulocytes, RBC, hemoglobulin, and hematocrit were counted. And plasma concentration of erythropoietin (EPO) which is the major regulator of erythropoiesis was measured using $\^$125/I-antierythropoietin IgG. Ferritin concentration in plasma was also analyzed.

(중략) 금번 조사한 양서류 이상의 고등동물에 있어서는 rat를 제외하고 Mt에 있어서의 26-수산화활성이 Ms에서 보다 높았으며, 그 입체 배치도 25R체가 우세하게 생성되었다. 종래, 이 동물종의 연구에 주되게 쓰여져 왔던 rat가 예외적인 존재였다. 한편, 어류에 있어서는 그 결과가 다른 동물종과 큰 차이가 있었다. 즉, 잉어는 Ms의 활성이 Mt에 비해 압도적으로 높았으며 25R체가 대부분 생성되었다. 도미는 어느 fraction에서도 25S체가 극미량밖에 생성되지 않았으며, 송어의 경우 26-수산화활성은 검출되지 않았다. 여기서 송어의 담즙주성분은 cholic acid이며, 이 동물의 경우에는 통상과는 다른 담즙산 생합성경로의 가능성이 시사되었다.

The glucuronide conjugates of morphine have been claimed to exert significant neuropharmacological effects. Morphine-6-glucuronide (M6G) may be a potent opioid agonist in vivo, and morphine-3-glucuronide (M3G) may act as a weak opioid antagonist. The present study addressed the permeability of the blood-brain barrier (BBB) for these metabolites compared to morphine. Tracers were prepared by enzymatic glucuronidation of U-methyl-$^3$H]-morphine. Brain uptake in rats was measured by the internal carotid artery perfusion technique and after i.v. bolus injections. In the perfusion experiments morphine showed a permeability-surface area product (PS) of 3.52${\pm}$0.61 ${\mu}$L min$\^$-1/ g$\^$-1/ Uptake seems to be mediated by passive diffusion and was not saturable by 100 ${\mu}$M morphine in the perfusate. The BBB permeability of [$^3$H]-M3G and [$^3$H]-M6G was too low to be quantified after 5 min of perfusion. Brain uptake of [$^3$H]-M3G and [$^3$H]-M6G 60 min after i.v. bolus injection reached 0.0060${\pm}$0.0003 and 0.0030${\pm}$0.0005% injected dose per g, respectively. From these brain concentrations and from the corresponding plasma concentration - time curves, BBB PS values of 0.14${\pm}$ 0.02 ${\mu}$L min$\^$-1/g$\^$-1/ and 0.11 ${\pm}$ 0.01 ${\mu}$L min$\^$-1/g$\^$-1/, respectively, were calculated. The ratio of BBB PS values is complementary to the analgesic potencies of morphine and M6G after different routes of administration. The low PS of MSG explains, why it is approximate]y equipotent to morphine after systemic injection, although it is about 2 orders of magnitude more potent than morphine after administration directly into the central nervous system.

Previous work in our laboratory has shown that noncompetitive N-methyl-D-aspartate (NMDA) receptor antagonists, MK-801, ketamine, dextrorphan and dextromethorphan cause a pronounced inhibition of apomorphine-induced cage climbing behavior in intact mice, suggesting the involvement of NMDA receptors in the glutamatergic modulation of dopaminergic function at the postsynaptic dopamine (DA) receptors: Therefore, in order to definitively establish the involvement of NMDA receptor in the apomorphine-induced dopaminergic response at the postsynaptic DA receptor, it is necessary to investigate whether or not the noncompetitive NMDA receptor antagonists would inhibit these phenomena not only in intact mice but also in the mice that are devoid of any involvement of indirect dopaminergic function. To minimize the risk of any indirect involvement of NMDA antagonists with DA neurons, vesicular DA stores were first depleted with reserpine.

경구투여급성독성시험 : 암수동물의 모든 투여군에서 시험물질투여에 의한 일반증상, 체중변화 및 육안적 부검소견은 관찰되지 않았으며. 따라서 반수치사량(LD$_{50}$)은 암수동물의 경우 5000mg/kg이상이었다. 경구투여아급성독성시험 : SD랫트의 암수 동물에 있어서 Captopril(SR Tablet 25mg)의 2000mg/kg 용량은 확실중독량에 해당되며, 무영향량(NOEL)은 220mg/kg용량이라고 사료되었다.

Gentamicin은 임상에서 많이 사용되는 aminoglycoside계 항생물질로서 세균의 세포막 단백질 합성을 억제하여 살균작용을 나타낸다. 최근 Gentamicin이 동물사료에 포함되거나 동물약품으로 많이 사용되어, 이를 복용한 식용가축에서의 잔류 량에 대한 인체유해성이 WHO/FAO 식품첨가물 전문가 협의회에서 논의되고 있다. Gentamicin의 육가공류의 잔류허용량 기준설정을 위한 독성 재평가의 일환으로 in vivo. in vitro 염색체이상시험을 실시하여 다음과 같은 결론을 얻었다. 1. 체외 염색체이상시험에서는 포유동물 배양세포인 chinese hamster lung cell을 배양하여 gentamicin sulfate 및 gentamicin을 최고 처리농도 5mg/$m\ell$부터 세포독성시험을 실시한 결과, 세포독성을 나타내지 않았다. 본 시험에서는 5mg/$m\ell$를 최고농도로 2.5, 1.25mg/$m\ell$의 3농도를 직접법 및 대사활성화법으로 각 농도당 2매의 플레이트씩 슬라이드를 제작, 결과를 판독한 결과, 직접법 및 대사활성화법 모두에서 전 농도 군에서 음성대조군과 같은 정도의 염색체이상을 유발하여 유전독성이 없음을 나타내었다. 2. 체내 염색체 이상시험에서는 ddY마우스를 이용하여 gentamicin sulfate의 LD$_{50}$의 1/2에 해당하는 200mg/kg을 최고농도로 gentamicin 과 gentamicin sulfate를 암수 각각 3마리씩 공비 2의 3농도로 투여한 후, 24시간째 골수세포의 염색체 표본을 제작하여 관찰한 결과, 세포독성 및 염색체 이상을 유발하지 않았다. 또한 동물약품으로 사용되는 치료용량 및 투약방법에 근거하여 10mg/kg 및 5, 2.5mg/kg을 1일 1회씩 4회 투여한 군에서도 암수에 상관없이 전 농도 군에서 염색체이상을 나타내지 않아 유전독성을 나타내지 않음을 관찰하였다.

Eye irritation, primary skin irritation and skin sensitization tests for Aloewhite were tested in New Zealand White rabbits and Hartley guinea pig. In primary skin irritation test of male New Zealand White rabbits, body weights were not significantly changed and primary Irritation Index(PII) was 0.47, indicating Aloewhite as mildly irritating material, In ocular irritation test, any injury on iris, conjunctival membrane, and cornea in New Zealand White rabbits was not observed. No injuries of the ocular mucous membrane were also recorded. Skin sensitization was tested in guinea Peg after intradermal and epicutaneous induction and graded I with zero % sensitization rate. These results indicate that Aloewhite was not considered to be irritant in test organs of animals.

-Acute oral and subcutaneous toxicity of Aloewhite(30% aloesine) were carried out in ICR mice. In this study, we daily examined number of deaths, clinical signs, body weights, and pathological examinations for 14 days after single oral and subcutaneous administration of Aloewhite with different dose levels. Aloewhite did not show any remarkable toxic effect in mice. These results suggest that oral and subcutaneous LD$\sub$50/ values in mice were over 6.8g/kg and 10g/kg, respectively.

We Investigated the peripheral excitatory effect of capsaicin and KR-25018, a newly synthesized capsaicin derivative which was demonstrated to have a potent analgesic activity. KR-25018 and capsaicin were found to be both potent efficacious contractors of isolated guinea pig bronchial smooth muscle. KR-25018 was equipotent with capsaicin and [Sar$\^$9/,Met(O$_2$)$\^$11/]-substance P, 10-fold more potent than histamine and 10-fold less potent than (${\beta}$ -Ala$\^$8/)-neurokinin A(4-10), and their -log(M)EC$\_$50/ values were 6.94${\pm}$0.08, 6.86${\pm}$0.05, 6.96${\pm}$0.07, 5.64${\pm}$0.04, 7.96${\pm}$0.02, respectively. Contractile responses to KR-25018 and capsaicin were potentiated by phosphoramidon (1 ${\mu}$M), an inhibitor of neuropeptide-inactivating endopeptidase, but completely abolished in a calcium-free medium. These responses to KR-25018 and capsaicin were unaffected by the NK-1 antagonist CP96345 (1${\mu}$M), partially inhibited by the NK-2 antagonist SR48968 (1 ${\mu}$M) but almost completely abolished by a combination of the antagonists. A vanilloid receptor antagonist capsazepine competitively antagonized the responses to both KR-25018 and capsaicin (pA$_2$: aganst KR-25018, 5.98${\pm}$0.47; against capsaicin, 5.80${\pm}$0.31), and a capsaicin-sensitive cation channel antagonist ruthenium red caused significant reduction in the maximum responses to KR-25018 and capsaicin (pD'$_2$: against KR-25018, 4.61${\pm}$0.33; against capsaicin 4.96${\pm}$0.21). In conclusion, the present results suggest that KR-25018 and cpasaicin act on the same vanilloid receptor inducing the influx of calcium through ruthenium red-sensitive cation channel and produce contractile responses via the release of tachykinins that act on both NK-1 and NK-2 receptor subtypes.

KR-25018 과 capsaicin 모두 용량의존적으로 진통효과를 나타내었으며 효력은 KR-25018이 capsaicin 보다 약간 강하거나 비슷하였고, 두 약물 모두 10 mg/kg에서 최대 효과를 나타내었다. KR-25018 과 capsaicin의 투여 농도에 의존적으로 기관지의 substance P 양이 감소하였으며 두 약물 모두 5 mg/kg에서 최대효과를 나타내었다. Capsaicin 1 mg/kg 전처치에 의해 기니픽의 체온이 30 분부터 감소하였으나, KR-25018 1 mg/kg에 의해서는 체온변화가 훨씬 미약하였다. Capsazepine 의 병용투여에 의해 두 약물의 진통작용 및 substance P 감소작용은 모두 억제되었으나 체온 변화에 대해서는 capsazepine이 아무런 영향을 미치지 않았다.

Eosinophils are known to be important effector cells in pathogenesis of asthma. The elucidation of mechanism by which eosinophil survival is regulated in vivo at sites of inflammation is critical tn our understanding of asthma pathogenesis. The maintenance of these cells at site of inflammation depends upon tile balance between its tendency to undergo apoptosis and tile local eosinophil-viability enhancing activity, Qualitative and quantative phenotypic differences have been observed between bronchoalveolar lavage (BAL) and peripheral blood (PB) eosinophils (EOS). We hypothesize that BAL EOS Possess altered functional feature compared to PB EOS. BAL and PB EOS were obtained from ragweed allergic asthmatics after segmental antigen challenge (SAC) at 24 hour or one week, and purified over percoll and CDl6 negative selection. Cells were cultured in duplicate in RPMI, 15% FCS and 1% penicillin/streptomycin without exogenous cytokines. Eosinophil purity and viability was >92%. BAL. EOS viability was 69${\pm}$4.4% versus 39${\pm}$1.6% for PB EOS (p<0.005) at 48 hour time point, and this difference was maintained through day 5 (32${\pm}$7.6% vs. 3.0${\pm}$ 1.4%, p<0.05), Among BAL EOS, those harvested one week after SAC appeared to have an prolonged survival compared to those harvested at 24 hour. Coculture of BAL and PB EOS resulted in significant viability enhancement than expecteed. Direct neutralization of GM-CSF activity, not IL-3 and EL-5, markedly decreased tile survival of BAL EOS in culture, and abrogated tile viability enhancing activity of their culture supernatants in a dose dependent manner. We conclude that BAL EOS activated in vivo possess enhanced viability compared to PB EOS. Mixing and neutralization experiments suggest a role for autocrine production of GM-CSF.

Flavonoids are the large class of plant-derived polyphenolic compounds with a broad spectrum of biological actions including anti-inflammatory, anti-allergic and anti-asthmatic activities. It has been reported that quercetin and some other flavonoids inhibit the histamine release from mast cells. Disodium cromoglycate, a kind of biflavonoid, have been used as a anti-asthmatic agent. Therefore, the following study was undertaken to characterize tile structure-activity relationships of flavonoids in the inhibition of histamine release, from IgE-sensitized mast cells. Flavonoid acetate were synthesized by addition of acetic anhydride to the flavonoids dissolved in pyridine. According In the Mota method, IgE-rich serum was made in rats sensitized with Pertussis vaccine and ovalbumin. Rat abdominal mast cells were harvested and purified over tile discontineous Percoll gradients, Mast cells were preincubated in RPMI with IgE-rich serum and further incubated with flavonoids. Histamine contents released from mast cells were determined fluorospectrophotometrically after stimulation with ovalbumin. (omitted)

The 116 annual meeting of the pharmaceutical Society of Japan was held in Kanazawa from March 27 to 29 in 1996. Our presentation will be concerned to a few topics in this meeting. Recently not only pharmaco and toxico kinetic studies but also pharmaco and toxico dynamic studies are beginning to be worthy of notice. This is quite reasonable. because the blood concentration time curves of a parent compound must be useful, but it is no more than a cross section for pharmaco and toxico dynamics in many organs and tissues as main reaction site for drug metabolism.

수용성 DDB유도체(비스알킬아미노메톡시 디메틸 -4,4'-디메톡시-5,5',6,6'-디메톡시 대옥시디페틸-2,2'-디카복실레이트)를 합성하게 됨으로서 생체이용효율을 극대화시킬 수 있고 치료효과가 확실한 정맥주사제로의 제제화가 가능하리라고 사료된다.

새로운 발모제 개발에 관한 연구의 일환으로 현재 혈압 강하 및 발모제로 쓰여지고 있는 미녹시딜에 신나밀 클로라이드를 반응시켜 N-4-cinnamylamido-2-amino-6-piperidinopyrimidine 3-oxide (이하 MINAMYL 이라 칭함)을 만들어 이 화합물에 대하여 액성영향 및 반응시간 등 제반물성을 검토하였으며, 또한 정량법 및 결합구조 등에 관하여 검토하였다.

발모제에 관한 연구의 일환으로 현재 발모제로 쓰여지고 있는 미녹시딜에 구충제 또는 모근 혈관확장제 둥의 효능을 갖고 있는 씬나믹산을 반응시켜 새로운 화합물 (이하 CINMI라 칭함)을 만들었다. 이 화합물에 대하여 액성영향 및 반응시간 등 제반물성을 검토하고, 최적조건에서 흡광도를 측정하여 검량선을 작성하였으며, 그리고 결합구조 및 육모효과 등에 대하여 검토하였다.

미녹시딜은 말초혈관확장제로서 중증의 고혈압치료에 사용하여 왔으나, 이 약물이 투여된 고혈압환자에게서 다모증 현상이 나타난 이후 탈모증의 치료제로서 연구개발되었다. 그러나 미녹시딜은 투약중지와 더불어 모발의 성장이 중지되며 치료전의 상태로 환원되는 기간이 짧은 단점이 있으므로 이를 개선하기 위해 미녹시딜의 4-NH$_2$ 위치에 의약품의 각막박리에 쓰여지고 있는 살리실산을 도입하여 두피 모근 세포로의 약물의 침투성을 높여, 보다 지속적인 새로운 발모제를 개발하고자 하였다.

PG의 농도가 AM의 경피투과에 미치는 영향은 용해도곡선과 유수분배곡선의 교점인 40% 농도에서 투과극대를 보였다. 지방산 및 지방산 알코올의 촉진효과는 리놀레인산＞올에인산＞올레일알코올＞ DHA=EPA=라우린산의 순이었고 PG에 리놀레인산 또는 올레인산을 첨가한 경우는 PG 단독에 비해 129.9 및 43.0배의 투과를 나타냈다. 또, 리놀레인산의 농도증가 (2,5,10%)에 따라 피부/매질 간의 분배계수가 가장 큰 5% 농도의 리놀레인산을 사용했을 때 투과효과가 최대로 나타났다. 한편, AM의 농도증가에 따른 투과효과에 있어서는 포화용해상태인 2% 약물농도에서 투과 flux가 가장 컸다. 라우릴황산타트륨의 첨가는 유의성 있는 효과를 발현하지 못했고 지방산과 병용시에도 지방산 단독 사용시에 비해 오히려 투과를 감소시켰다. 종차에 따른 AM의 피부투과에 있어서는 리놀레인산을 투과촉진제로 사용한 결과 흰주와 무모마우스의 투과 flux가 각각 23.78 및 213.29 $\mu\textrm{g}$/$\textrm{cm}^2$/hr로 무모마우스가 8.97배 높았다. 흰주의 추출액 및 호모지네이트 중에서의 AM의 대사에 있어서는 겉보기 1차 분해속도정수가 표피측추출액, 장막측추출액, 호모지네이트에서 각각 0.0105, 0.572 및 6.153$hr^{-1}$로 호모지네이트 중의 분해속도가 가장 빨라 AM은 피부투과 중에 가장 많이 분해됨을 알았다.

While ranitidine is well known to be absorbed rapidly, the underlying cause of variable bioavailability in intra- and inter-subjects has not been clarified yet. Intestinal permeability is a key controlling factor for oral absorption of highly soluble drugs, In the present study, intestinal ferfusions have been conducted to determine the intestinal permeabilities(Peffs) of ranitidine in the rats, dogs and humans and compared to the estimated fractions of dose absorbed (FAs) in humans. A new in vivo methodology, using a regional segmental perfusion technique, has been used in the dogs and humans. In situ single-pass perfusion experiments have been performed in the rats. In the dog and human studies, perfusion experiments have been conducted on two periods to determine the intrasubject variability, There was low significant intrasubject variation as compared to intersubject variation. The Peffs of ranitidine were 33%, 51%, and 45% inthe rats, dogs and humans, respectively. The FAs were approximately the same for all three species models, suggesting rats and dogs are good animal models for estimating the oral absorption of ranitidine in humans. In addition, the estimated extent of absorption of this drug is consistent with the average bioavailability, indicating that ranitidine has permeability-limited absorption characteristics. Supported by FDA Grant FD01462.

Recently a novel in vivo approach in dogs, using a regional segmental intestinal perfusion technique, has been developed. The perfusion tube consists of a highly sophisticated multichannel tube with two inflatable occluding balloons, which are placed in 10cm apart. The tube was introduced orally from the stomach through the upper jejunum under the guidance of solid-state pH meter. In the present study, four healthy dogs were infused in the proximal jejunum on two periods. The two perfusion experiments used the same flow rate, 2 $m\ell$/min, and the same perfusion solution to determine the intrasubject variability. The mean (${\pm}$ S. E.) fractions of ranitidine absorbed calculated from the perfusion data were 21.32${\pm}$2.01% (n=3) (1st period), 27.88 ${\pm}$ 17.54% (n=4) (2nd period), respectively. The effective permeabilities (Peffs${\times}$10$\^$4/) of ranitidine were 1.51${\pm}$0.47cm/sec (n=3) (1st period), 1.50 ${\pm}$ 0.31 cm/sec (n=4) (2nd period), respectively. The pH and osmolarity of perfusion solution were 7.50 ${\pm}$ 0.03 and 300 ${\pm}$ 0.06 mOsm/L, There was no significant intrasubject variation. Mixing equilibrium (steady-state) was reached at about 50 min. l-Phenylalanine was absorbed almost completely. Intrinsic intestinal wall permeability of ranitidine showed low permeable characteristics, suggesting permeability-limited absorption. The absorption of 1-phenylalanine, an actively transported nutrient, was not inhibited by ranitidine. The low intestinal membrane permeability is one of the important factors responsible for the variable oral absorption of ranitidine. Supported by FDA Grant FD01462-04 and KOSEF Grant.

1차적인 제형의 조제를 위해서 비이온성 계면활성제인 poloxamer 류, Tween 류, octylphenylether와 양심 계면활성제인betaine 류와 방부제인 croquat 류에 대한 시험을 실시하여 자극을 나타내지 않은 성분을 선별하였다. 이를 근거로 isotonic sodium borate 용액을 완충액으로 사용하여, 최종제형을 조제하여 시험한 결과 자극반응을 나타내지 알았으며, 본 제형을 실온과 4$0^{\circ}C$에서 2, 4, 6개월간 안정성 시험을 거친 후 시험시에도 전혀 자극반응을 나타내지 않았다.

Rudko method에 의한 단백질 제거능 시험 결과, 기존에 시판 중인 렌즈 세척제 보다 신규 처방화 된 렌즈 세척제의 단백질 제거능이 1.5배 이상 우수하였으며, 전자 주사 현미경 사진과 비교시 동일한 결과를 보임을 알 수가 있다.

합성한 BR-8702-2의 시험결과 원소분석에서는 이론치와 실험치가 잘 일치하였고, 구조확인을 위한 FT-IR 및 FT-NMR(H$^1$)등의 시험에서도 구조와 일치하는 결과치를 나타내었다. 합성물질의 정량법을 HPLC 및 구성원소인 P와 N을 이용한 정량법에서도 유의성있는 결과를 얻을 수 있었다. 또한 분말상태인 이 물질의 안정성은 고온 고습하에서도 양호하였으며, 각종 완충수용액에서 pH가 분해에 미치는 영향을 pH-rate profile로 나타낸 결과 pH7, 8, 9에서 안정하였고 pH8에서 k=1.3$\times$$10^{-4}$day으로 가장 안정하여 shelf life는 807일이었다. 광에 대해서도 k=5.8$\times$$10^{-4}$day로 매우 안정하였다.

Anti-CALLA Fab-RTA immunotoxins were constructed using two crosslinking agent, SMPT and SMCC, to generate a disulfide and a thioether bridge between Fab fragment of K269-65 MoAb and RTA toxin moieties, respectively. These immunotoxins were selectively immunoreative with CALLA$\^$+/ B-lineage Daudi cells. SMPT and SMCC mediated RTA immunotoxins were prepared with 49% and 53% of the RTA conjugation yields, respectively. SDS-PAGE analysis show that immunotoxins were constructed with major Fab-1 RTA (76kda), minor Fab-2RTA (106kda) and Fab-3RTA (136kda) compositions. The breakdown rates of immunotoxins were determined in the presence of glutathione by measuring the amount of reduced immunotoxins using size-exclusion HPLC. The SMCC immunotoxins were more resistant to the glutathione than SMPT immunotoxins. But, our data showed that the SMPT mediated disulfide bonded immunotoxins were much more active than the SMCC mediated thioether bonded immunotoxins to kill the target cells in vitro.

Judging from hydrocortisone concentration in dosing area, the extent of absorption was reduced in the liposome-gel formulation. However, higher and sustained skin concentrations of hydrocortisone were achieved for the liposome-gel as compared to the ointment. Drug concentration in both viable and deep skin reached its maximum within 0.5 h after application of both formulations to both skin types. Drug concentrations in both skins from the ointment declined with time, while those from the liposome-gel were greatly sustained. The sustainment by the liposome-gel was more remarkable in the viable skin than in the deep skin. Drug concentration in the viable skin could be maintained at a nearly constant level for over 8 h by applying the liposome-gel. As a result, a 5-fold higher viable skin drug concentration was obtained from the liposome-gel than from the ointment at 8 h after the application to the SC-removed skin. However, the plasma concentration of hydrocortisone at 4 h from the liposome-gel was only one-fourth (p<0.01) the value from the ointment when the drug was applied to the SC-removed skin, consistent with. the lower urinary (one-third, p<0.05) and fecal (one-half, p＜0.05) excretion. Conclusions : Retarded diffusion of the drug from the skin to the systemic blood stream appears to be a potential factor in the sustained skin concentration of hydrocortisone from the liposome-gel, Interaction of hydrocortisone in the skin with phosphatidylcholine, a component of the liposomes and skin, may well be a factor in retarding the diffusion of the drug in the skin.

Sudan IV는 2시간 관류시키는 동안 약 30%의 농도감소를 보였다 Sudan IV자체는 물에 녹지않고 리포솜에 의해서 가용화 되는 물질이기 때문에 관류액중에서 리포솜으로부터 release out되지 않는다. 따라서 situation I의 가능성은 희박하며 또 그 자체로는 물에 거의 녹지않아 흡수되지 않을 것으로 예상되는 물질이므로 Sudan IV의 관류액으로부터의 소실은 곧바로 situation II를 반영할 것이다. 또 리포솜의 구성물질인 PC역시 관류액중에서 농도감소를 나타내었는데 이로부터 난용성약물을 봉입한 리포솜이 어떻게 장관으로 흡수되는 지 예상할 수 있었다.

carbamazepine은 대표적인 항전간제로써, 약물의 유효 혈중 농도 범위가 좁아서 TDM(Theapeutic Drug Monitoring)을 시행하는 약물이다. 그리고 이 약물은 parent drug 뿐만 아니라 대사체의 하나인 carbamazepine-10,11-epoxide 역시 carbamazepine와 동일한 효과가 있는 것으로 알려져 있어, 임상적으로 TDM 시행시에는 carbamazepine의 carbamazepine-10,11-epoxide로의 대사에 소요되는 시간과 대사 정도에 대한 자료가 필요하다. 그러나 이제껏 제시되고 있는 population parameter들이 모두 서양인에 대한 자료이므로 국내에서 이 약물을 투여하는데 있어서 인종간의 차이를 확인하지 않고서 서양인의 자료에 준하여 적용하는 것은 상당한 위험성이 따를 수도 있으므로 한국인에 있어서의 carbamazepine 대사에 관한 연구가 필요하였다. 방법 7명의 건강한 성인을 대상으로 carbamazepine 제제 400mg을 1회 경구 투여한 다음, 0, 0.5, 1, 2, 3, 4, 6, 8, 12, 24, 36, 48, 72 시간 경과시 채혈하여 정량하였다. 결과 carbamazepine의 AUC 881$\pm$233(minㆍ$\mu\textrm{g}$/$m\ell$), MRT 72.1$\pm$10.8(min), t$_{1}$2/ 40.1 $\pm$ 8.6(min), CL 6.75 $\pm$ 2.72($m\ell$/min/kg), Vdssn 484 $\pm$ 215($m\ell$/kg)의 값을 얻었다.

The immunoassay method is frequently used for the identification and quantitation of ultratrace amount of bioactive substances. Homogeneous liposome immunoassays, which can avoid the use of radioisotopes and separation steps, have recently been reported in many publications. Cytolysin-mediated liposome immunoassay using melittin ever been studied but showed limited applications. Here, we designed a homogeneous liposome immunoassay using Clostridium perfringens phospholipase C (PLC), an enzyme which catalyzes the hydrolysis of phosphatidylcholine in biological membranes, as a cytolysin.

The purpose of this study was to prepare the nifedipine dry elixir (NDE) and coated nifedipine dry elixir (CNDE) containing nifedipine ethanol solution for improving the dissolution rate and bioavailability of nifedipine. NDE containing nifedipine and ethanol in wall materials of dextrin was prepared using a spray-dryer and then NDE was coated with eudragit acrylic resin to make CNDE. Shape and size of the NDE and CNDE were monitored by scanning electron micrograph and laser particle size analyzer In vitro dissolution tests were performed in simulated gastric and intestinal fluid. Bioavailability of NDE and CNDE were compared with drug powder suspension and commercial soft capsule after oral administration of the preparations to rats. NDE and CNDE are spherical in shape. Cross-sectional view of dry elixirs indicates the large inter cavity containing ethanolic drug solution in shell. Geometric mean diameter of NDE and CNDE is about 6.64 and 8.70 $\mu\textrm{m}$, respectively. Drug dissolution rate within first 5 min from NDE increased dramatically irrespective of dissolution medium. However, CNDE showed a particularly retarded dissolution rate in pH 1.2 simulated gastric fluid compared with NDE. The bioavailability of nifedipine in the NDE was increased dramatically compared with drug powder suspension. CNDE reduced initial burst-out plasma peak compared with NDE. CNDE as a sustained release delivery system could reduce the initial burst-out plasma peak due to controlling the release rate of nifedipine from NDE and maintain the effective plasma level over a longer period within therapeutic window with enhanced bioavailability of nifedipine.

The hydrolyzed-lactose milk for the lactase-deficient subject is sweeter than whole milk, and some subjects dislike its taste. To overcome this shortcoming the dried liposomes containing ${\beta}$-galactosidase to digest lactose in milk after drinking were prepared and examined the possible application of this dried liposomes to the lactase-deficient subjects. To improve the stability of conventional liposome suspension, the dried liposomes in the presence of trehalose were prepared by the dehydration-rehydration vesicles method. Small unilamellar vesicles, prepared with egg phosphatidyl cholesterol, and cholesterol, were mixed with ${\beta}$-galactosidase solution and then ;up[jo;ozed. The freeze-dried liposome was rehydrated and centrifuged. The resultant multilamellar vesicles were mixed with trehalose(4g/g lipid) and then lyophilized to produce final dried liposome. Trehalose increased the entrapping efficiency of liposomes by 3 fo1d compared to the liposomes without trehalose (13% vs. 46%).

Titrated Extract of Centella asiatica (TECA) is a poorly water-soluble extract from the Centella asiatica. Despite excellent wound preparation causes pain due to a low aqueous solubility and high hypertonicity and therefore, the patient's compliance of this drug is low. The objective of this study is to design a formulation of TECA with an improved therapeutic applicability via an adequate solubilization. A mixture of propylene glycol and ethoxylated hydrogenated caster oil achieved an acceptable solubilization of TECA (i.e. 10%) via a formulation of micelle. A preparation of extemporaneous TECA micelle was achieved by a dilution of the original micellar formulation with either water or saline. An estimated distribution of particle size was between 15.9 and 32.6 ㎜. The osmotic pressure of the formulation was found to be much lower than that found In a commercially available injectable (i.e. Madecassole). The erthrocytic hemolysis of micellar solution was lower than that with the conventional dosage form, consistent with the osmotic pressure data. Pain score after an injection of the micellar solution was assessed by the hind-paw writhing test using ICR mice and compared with that found with the conventional injectable. The data indicated that the injection of the micellar solution was a significantly less painful. These results indicated that a micellar solubilization, followed by an extemporaneous dilution, is a novel formulation of TECA with an improved therapeutic applicability.

Cyclosporin A (CyA) is widely used in the inhibition of graft rejection in organ transplantation. However, the bioavailability of CyA after oral administration is very low due to its poor solubility and dispersability hi water. To improve the solubility of CyA, microemulsion systems were developed and its physicochemical characteristics were evaluated by phase studies, solubility and dispersability tests. Phase studies on the systems composed of ethyl oleate (EO), PPG-20 methyl glucose ether (GP-20), poloxamer 123 (PL) and water U) were carried out to make stable w/o emulsions. Besides, based on CyA solubility test in various compositions of surfactant systems, a reasonable surfactant composition (GP-20/PL=4/1) was selected to enhance its solubility.

In order to elucidate the mechanism of action of transdermal absorption-enhancing compounds, i.e., pyrrolidone derivatives (2-pyrrolidone, 1-methyl-2-pyrrolidone, 1-ethyl-2-pyrrolidone, 1,5-dimethyl-pyrrolidone and 5-methyl-2-pyrrolidone), multilamellar liposome was prepared from the simulated stratum corneum lipid and employed as a model system for the barrier function of the stratum corneum. The liposomal membrane of the stratum corneum lipid liposome (SCLL) behaves as an osmometer and has an excellent barrier function. In addition, its phase transition temperatures are similar to those of human stratum corneum intercellular lipid region. Therefore, SCLL seems to be a useful skin model. To estimate the barrier function of SCLL, the osmotic behavior of SCLL was measured in the presence of pyrrolidone derivatives and the effect on the phase transition temperature of SCLL was also investigated using differential calorimetry. Above a certain concentration (MLAC), enhances perturb the barrier function of the liposome. The relationship between MLACs and the partition coefficient of the pyrrolidone derivatives was observed; the greater the partition coefficients, the smaller the MLAC. This suggests that the more hydrophobic enhancers penetrate into the lipid layer more easily and reduce the barrier function of membrane more effectively. The results of differential scanning thermograms of the SCLL suggest that the pyrrolidone derivatives had incorporated into the lipid layer in the liposome and increased the fluidity of the lipid layer in the liposome. Such activity might have some correlation with the transdermal absorption-enhancing activity these compounds.

Long-circulating liposomes have prompted interests in using them as a drug delivery system. This improvements in delivery has been thought to be the results from sustained action of liposomes in plasma without RES uptake. Although methotrexate(MTX) has been one of the most widely used antineoplastc drug, its use was limited by prompt RES uptake. The purpose of this study was to prepare long-circulating liposomes for MTX using highly water-soluble polymer (PEG-PE). In vitro, release of MTX from liposomes in phosphate buffer (pH 7.4), rat liver homogenate, and rat plasma was investigated. The pharmacokinetics and organ distribution of fee drug, conventional and long-circulating liposomes were also compared with one another after intravenous administration to rats.

The microcapsules containing ethanol in a water-soluble polymer shell could be formed by the spray-drying technique. A mixed solution of ethanol, water and a water-soluble polymer is spray-dried at the lowest possible temperature to obtain a powder product in which the water is substantially removed and the ethanol is encapsulated in water-soluble polymer shell because of the hydrophillic property of polymer and permeability difference between ethanol and water. In this study, the effect of sodium lauryl sulfate (SLS) on the ethanol content, ethanol encapsulation efficiency and yield were investigated to maximize the microencapsulation of ethanol.